Methods for treating cancers using oral formulations of cytidine analogs

ABSTRACT

Provided herein are methods for treating subjects having a cancer, such as a relapsed or refractory solid tumor, wherein the method comprises administering to the subject a cytidine analog. In certain of the methods, the cytidine analog is administered alone or in combination with one or more anti-cancer agents. Also provided are methods for using a cytidine analog, to treat diseases and disorders including disorders related to abnormal cell proliferation, hematologic disorders, and immune disorders, among others. In certain of the methods, the cytidine analog is formulated in an oral dosage form and administered orally.

This application claims priority to, and is a continuation-in-part ofU.S. non-provisional application 13/665,778, filed Oct. 31, 2012, whichclaims priority to U.S. Provisional Patent Application No. 61/554,344,filed Nov. 1, 2011, both of which are hereby incorporated by referencein their entirety.

I. FIELD

Provided herein are methods for treating, preventing, or managingcancers using a cytidine analog, or a salt, solvate, or hydrate thereof.Also provided are methods for using a cytidine analog, or a salt,solvate, or hydrate thereof, to treat, prevent, or manage diseases anddisorders including disorders related to abnormal cell proliferation,hematologic disorders, and immune disorders, among others. In certain ofthe methods, the cytidine analog is formulated in an oral dosage formand administered orally.

II. BACKGROUND

Cancer is a major worldwide public health problem; in the United Statesalone, approximately 570,000 cancer-related deaths were expected in2005. See, e.g., Jemal et al., CA Cancer J. Clin. 55(1):10-30 (2005).Many types of cancer have been described in the medical literature.Examples include cancer of the blood, bone, lung (e.g., non-small-celllung cancer and small-cell lung cancer), colon, breast, prostate, ovary,brain, and intestine. The incidence of cancer continues to climb as thegeneral population ages and as new forms of cancer develop. A continuingneed exists for effective therapies to treat subjects with cancer.

Nucleoside analogs have been used clinically for the treatment of viralinfections and certain cancers. Most nucleoside analogs are classifiedas anti-metabolites. After they enter the cell, nucleoside analogs aresuccessively phosphorylated to nucleoside 5′-mono-phosphates,di-phosphates, and tri-phosphates.

The nucleoside analogs 5-azacytidine (also known as4-amino-1-β-D-ribofuranosyl-1,3,5-triazin-2(1H)-one; National ServiceCenter designation NSC-102816; CAS Registry Number 320-67-2;azacitidine; Aza and AZA; and currently marketed as VIDAZA®) and2′-deoxy-5-azacytidine (also known as 5-aza-2′-deoxycytidine,decitabine, 5-aza-CdR, Dac, and DAC, and currently marketed as DACOGEN®)are DNA methyltransferase (DNMT) inhibitors that have been approved bythe U.S. Food and Drug Administration for the treatment ofmyelodysplastic syndromes (MDS). Azacitidine and decitabine are cytidineanalogs; a structural difference between these cytidine analogs andtheir related natural nucleosides is the presence of a nitrogen atposition 5 of the cytosine ring in place of a carbon. Azacitidine may bedefined as having a molecular formula of C₈H₁₂N₄O₅, a molecular weightof 244.21 grams per mole, and a structure as shown below. Decitabine maybe defined as having a molecular formula of C₈H₁₂N₄O₄, a molecularweight of 228.21 grams per mole, and a structure as shown below.

After its incorporation into replicating DNA, 5-azacytidine or5-aza-2′-deoxycytidine can form a covalent complex with DNAmethyltransferases. DNA methyltransferases are responsible for de novoDNA methylation and for reproducing established methylation patterns indaughter DNA strands of replicating DNA. Inhibition of DNAmethyltransferases can lead to DNA hypomethylation, thereby restoringnormal functions to morphologically dysplastic, immature cells byre-expression of genes involved in normal cell cycle regulation,differentiation and death. The cytotoxic effects of cytidine analogs cancause the death of rapidly dividing cells that are no longer responsiveto normal cell growth control mechanisms. 5-Azacytidine, unlike5-aza-2′-deoxycytidine, also incorporates into RNA. The cytotoxiceffects of azacitidine may result from multiple mechanisms, includinginhibition of DNA, RNA and protein synthesis, incorporation into RNA andDNA, and activation of DNA damage pathways.

5-Azacytidine and 5-aza-2′-deoxycytidine have been tested in clinicaltrials and showed significant activity, such as, for example, in thetreatment of myelodysplastic syndromes (MDS), acute myelogenous leukemia(AML), chronic myelogenous leukemia (CML), acute lymphocytic leukemia(ALL), and non Hodgkin's lymphoma (NHL). See, e.g., Aparicio et al.,Curr. Opin. Invest. Drugs 3(4): 627-33 (2002). 5-Azacytidine hasundergone NCI-sponsored trials for the treatment of MDS and has beenapproved for treating all FAB subtypes of MDS. See, e.g., Kornblith etal., J. Clin. Oncol. 20(10): 2441-2452 (2002); Silverman et al., J.Clin. Oncol. 20(10): 2429-2440 (2002). 5-Azacytidine may alter thenatural course of MDS by diminishing the transformation to AML throughits cytotoxic activity and its inhibition of DNA methyltransferase. In aPhase III study, 5-azacytidine administered subcutaneously significantlyprolonged survival and time to AML transformation or death in subjectswith higher-risk MDS. See, e.g., P. Fenaux et al., Lancet Oncol., 2009,10(3):223-32; Silverman et al., Blood 106(11): Abstract 2526 (2005).

Other members of the class of cytidine analogs include, for example:1-β-D-arabinofuranosylcytosine (Cytarabine or ara-C); pseudoisocytidine(psi ICR); 5-fluoro-2′-deoxycytidine (FCdR);2′-deoxy-2′,2′-difluorocytidine (Gemcitabine);5-aza-2′-deoxy-2′,2′-difluorocytidine; 5-aza-2′-deoxy-2′-fluorocytidine;1-β-D-ribofuranosyl-2(1H)-pyrimidinone (Zebularine);2′,3′-dideoxy-5-fluoro-3′-thiacytidine (Emtriva); 2′-cyclocytidine(Ancitabine); 1-β-D-arabinofuranosyl-5-azacytosine (Fazarabine orara-AC); 6-azacytidine (6-aza-CR); 5,6-dihydro-5-azacytidine(dH-aza-CR); N⁴-pentyloxycarbonyl-5′-deoxy-5-fluorocytidine(Capecitabine); N⁴-octadecyl-cytarabine; and elaidic acid cytarabine.

5-Azacytidine and certain other cytidine analogs are approved forsubcutaneous (SC) or intravenous (IV) administration to treat certainproliferative disorders. Oral dosing of cytidine analogs would be moredesirable and convenient for patients and doctors, e.g., by eliminatinginjection-site reactions that may occur with SC administration and/or bypermitting improved patient compliance. However, oral delivery ofcytidine analogs has proven difficult due to combinations of chemicalinstability, enzymatic instability, and/or poor permeability. Forexample, cytidine analogs have been considered acid labile and unstablein the acidic gastric environment. Previous attempts to develop oraldosage forms of cytidine analogs have required enteric coating of thedrug core to protect the active pharmaceutical ingredient (API) fromwhat was understood and accepted to be therapeutically unacceptablehydrolysis in the stomach, such that the drug is preferably absorbed inspecific regions of the lower gastrointestinal tract, such as thejejunum in the small intestine. See, e.g., Sands, et al., U.S. PatentPublication No. 2004/0162263 (application Ser. No. 10/698,983). Inaddition, a generally accepted belief in the art has been that waterleads to detrimental hydrolytic degradation of cytidine analogs duringformulation, subsequently affecting the stability of the API in thedosage form. As a result, coatings applied to the drug core forprospective oral delivery of cytidine analogs have previously beenlimited to organic solvent-based systems to minimize exposure of the APIto water.

III. SUMMARY

Provided herein are methods for treating, preventing, or managingcancers using a cytidine analog, or a salt, solvate, or hydrate thereof.Also provided are methods for using a cytidine analog, or a salt,solvate, or hydrate thereof, to treat, prevent, or manage diseases anddisorders, including disorders related to abnormal cell proliferation,hematologic disorders, and immune disorders, among others. In oneembodiment, the cancer is a solid tumor. In one embodiment, the canceris relapsed or refractory. In one embodiment, the cancer is a cancer ofthe breast, lung, head and neck, ovary, testicle, prostate,gastrointestinal system, stomach, pancreas, liver, colon, kidney,bladder, brain, skin, or bone, among others. In one embodiment, thecancer is a cancer of the blood or the lymph. In particular embodiments,the cancer is a relapsed or refractory solid tumor. In particularembodiments, the cancer is a cancer of the bladder, ovary, pancreas,lung, colon, head and neck, breast, or skin. In particular embodiments,the cancer is a cancer of the bladder, ovary, pancreas, lung, or colon.

In one embodiment, the cytidine analog is formulated in an oral dosageform provided herein (e.g., a tablet or a capsule). In one embodiment,the cytidine analog is administered orally to a subject in need thereof.In one embodiment, the cytidine analog is administered to a subject inneed thereof for a sustained period of time. In one embodiment, thecytidine analog is administered to a subject in need thereof cyclically(e.g., dosing for one or more days, followed by a resting period). Inone embodiment, the cytidine analog is administered to a subject in needthereof over multiple dosing cycles.

In one embodiment, the cytidine analog is administered alone as a singleagent to a subject in need thereof. In one embodiment, the cytidineanalog is administered in combination with one or more additionalanti-cancer agent(s), including, but not limited to, carboplatin,paclitaxel, or Abraxane® (paclitaxel protein-bound particles), amongothers. In one embodiment, the additional anti-cancer agent is analkylating agent, a cytotoxic agent, an anti-angiogenic agent, ananti-tubulin agent, an anti-metabolite, a kinase inhibitor, a biologicsagent, or any other known anti-cancer agent (e.g., an anti-cancer agentprovided herein elsewhere). In certain embodiments, in addition to thecytidine analog or the one or more additional anti-cancer agent(s), ananti-emetic is administered to a subject in need thereof.

In one embodiment, the cytidine analog is administered orally orparenterally. In one embodiment, the cytidine analog is administeredorally. In particular embodiments, 5-azacytidine is administered orally.In one embodiment, the additional anti-cancer agent is administeredorally or parenterally. In one embodiment, the cytidine analog isadministered via the same route as the one or more additionalanti-cancer agent(s). In one embodiment, the cytidine analog isadministered via a different route as the one or more additionalanti-cancer agent(s) (e.g., one administered orally and the otheradministered parenterally).

In one embodiment, the cytidine analog is administered in a particulardosing cycle. In one embodiment, the cytidine analog and the one or moreadditional anti-cancer agent(s) (including, but not limited to,carboplatin, paclitaxel, or Abraxane®) are co-administered in aparticular dosing cycle. In particular embodiments, the cytidine analogis first administered to a subject in need thereof for one or more days(e.g., for 7 days or more), and the one or more additional anti-canceragent(s) is/are administered to the subject (e.g., starting on Day 8 orlater of the treatment cycle). In particular embodiments, when the oneor more additional anti-cancer agent(s) is/are administered to thesubject, the cytidine analog is also administered to the subject. Inparticular embodiments, when the one or more additional anti-canceragent(s) is/are administered to the subject, the cytidine analog is notadministered to the subject simultaneously.

In one embodiment, provided herein are pharmaceutical compositionscomprising a cytidine analog, wherein the compositions release the APIsubstantially in the stomach upon oral administration. In oneembodiment, provided herein are pharmaceutical compositions comprising acytidine analog, wherein the compositions release the API substantiallyin the stomach and the upper intestine upon oral administration. Alsoprovided are methods for making the compositions, and methods for usingthe compositions to treat, prevent, or manage diseases and disordersincluding cancer, disorders related to abnormal cell proliferation,solid tumors, and hematologic disorders.

In certain embodiments, the cytidine analog is 5-azacytidine. In otherembodiments, the cytidine analog is 5-aza-2′-deoxycytidine (decitabineor 5-aza-CdR). In yet other embodiments, the cytidine analog is, forexample: 1-β-D-arabinofuranosylcytosine (Cytarabine or ara-C);pseudoisocytidine (psi ICR); 5-fluoro-2′-deoxycytidine (FCdR);2′-deoxy-2′,2′-difluorocytidine (Gemcitabine);5-aza-2′-deoxy-2′,2′-difluorocytidine; 5-aza-2′-deoxy-2′-fluorocytidine;1-β-D-ribofuranosyl-2(1H)-pyrimidinone (Zebularine);2′,3′-dideoxy-5-fluoro-3′-thiacytidine (Emtriva); 2′-cyclocytidine(Ancitabine); 1-β-D-arabinofuranosyl-5-azacytosine (Fazarabine orara-AC); 6-azacytidine (6-aza-CR); 5,6-dihydro-5-azacytidine(dH-aza-CR); N⁴-pentyloxycarbonyl-5′-deoxy-5-fluorocytidine(Capecitabine); N⁴-octadecyl-cytarabine; elaidic acid cytarabine; ortheir derivatives or related analogs.

Certain embodiments herein provide compositions that are single unitdosage forms comprising a cytidine analog. Certain embodiments hereinprovide compositions that are non-enteric-coated. Certain embodimentsherein provide compositions that are tablets comprising a cytidineanalog. Certain embodiments herein provide compositions that arecapsules comprising a cytidine analog. In certain embodiments, thesingle unit dosage forms optionally further contain one or moreexcipients. In certain embodiments, the tablets optionally furthercontain one or more excipients. In other embodiments, the capsulesoptionally further contain one or more excipients. In certainembodiments, the composition is a tablet that effects an immediaterelease of the API upon oral administration. In other embodiments, thecomposition is a tablet that effects a controlled release of the APIsubstantially in the stomach. In other embodiments, the composition is atablet that effects a controlled release of the API substantially in thestomach and the upper intestine. In certain embodiments, the compositionis a capsule that effects an immediate release of the API upon oraladministration. In other embodiments, the composition is a capsule thateffects a controlled release of the API substantially in the stomach. Inother embodiments, the composition is a capsule that effects acontrolled release of the API substantially in the stomach and the upperintestine. In particular embodiments, the tablet contains a drug corethat comprises a cytidine analog, and optionally further contains acoating of the drug core, wherein the coating is applied to the drugcore using an aqueous solvent, such as, for example, water, ornon-aqueous solvent, such as, for example ethanol.

Certain embodiments herein provide methods of making formulations ofcytidine analogs intended for oral delivery. Further provided arearticles of manufacture containing packaging material, an oralformulation of a cytidine analog, and a label that indicates that theformulation is for the treatment, prevention, or management of certaindiseases or disorders including, e.g., a cancer, a disorder related toabnormal cell proliferation, a solid tumor, a hematologic disorder, oran immune disorder.

Certain embodiments herein provide methods of using the formulationsprovided herein to treat, prevent, or manage diseases or disordersincluding, e.g., cancer, disorders related to abnormal cellproliferation, solid tumors, hematologic disorders, or immune disorders.In certain embodiments, the formulations of cytidine analogs are orallyadministered to subjects in need thereof to treat, prevent, or manage acancer; or a hematological disorder, such as, for example, MDS, AML,ALL, CML, NHL, leukemia, lymphoma, or multiple myeloma; or a solidtumor, such as, for example, sarcoma, melanoma, carcinoma, or cancer ofthe colon, breast, ovary, gastrointestinal system, kidney, bladder, lung(e.g., non-small-cell lung cancer and small-cell lung cancer), testicle,prostate, stomach, pancreas, liver, head and neck, brain, skin, or bone,among others. In particular embodiments, the cancer is a cancer of thebladder, ovary, pancreas, lung, colon, head and neck, breast, or skin.In particular embodiments, the cancer is a cancer of the bladder, ovary,pancreas, lung, or colon. In certain embodiments, the cancer isrefractory. In certain embodiments, the cancer is relapsed. In certainembodiments, the cancer is metastatic. In certain embodiments, theformulations of cytidine analogs are orally administered to subjects inneed thereof to treat, prevent, or manage an immune disorder. In certainembodiments, the oral formulations provided herein are co-administeredwith one or more therapeutic agents to provide a synergistic therapeuticeffect in subjects in need thereof. In certain embodiments, the oralformulations provided herein are co-administered with one or moretherapeutic agents to provide a resensitization effect in subjects inneed thereof. The co-administered agents may be a cancer therapeuticagent, as described herein. In certain embodiments, the co-administeredagent(s) may be dosed, e.g., orally or by injection. In certainembodiments, the cytidine and/or the co-administered agent(s) may bedosed cyclically.

In particular embodiments, provided herein are tablets containing5-azacytidine and methods for making and using the tablets to treat,prevent, or manage cancer, disorders related to abnormal cellproliferation, solid tumors, or hematologic disorders. In certainembodiments, the tablets optionally further contain one or moreexcipients such as, for example, glidants, diluents, lubricants,colorants, disintegrants, granulating agents, binding agents, polymers,and/or coating agents. Examples of ingredients useful in preparingcertain formulations provided herein are described in, e.g., Etter etal., U.S. Patent Publication No. 2008/0057086 (application Ser. No.11/849,958), and Etter et al., U.S. Patent Publication No. 2009/0286752(application Ser. No. 12/466,213), both of which are incorporated hereinby reference in their entireties.

Specific embodiments herein provide, inter alia, pharmaceuticalcompositions comprising a therapeutically effective amount of5-azacytidine. Specific embodiments herein provide, inter alia,pharmaceutical compositions comprising a therapeutically effectiveamount of 5-azacytidine, wherein the composition releases the5-azacytidine substantially in the stomach following oral administrationto a subject. Further embodiments provide the aforementionedcompositions, which: are immediate release compositions; do not have anenteric coating (i.e., are non-enteric-coated); are tablets; arecapsules; further comprise an excipient selected from any excipientdisclosed herein; further comprise a permeation enhancer; furthercomprise d-alpha-tocopheryl polyethylene glycol 1000 succinate; furthercomprise a permeation enhancer in the formulation at about 2% by weightrelative to the total weight of the formulation; are essentially free ofa cytidine deaminase inhibitor; are essentially free oftetrahydrouridine; have an amount of 5-azacytidine of at least about 40mg; have an amount of 5-azacytidine of at least about 50 mg; have anamount of 5-azacytidine of at least about 60 mg; have an amount of5-azacytidine of at least about 80 mg; have an amount of 5-azacytidineof at least about 100 mg; have an amount of 5-azacytidine of at leastabout 120 mg; have an amount of 5-azacytidine of at least about 150 mg;have an amount of 5-azacytidine of at least about 200 mg; have an amountof 5-azacytidine of at least about 250 mg; have an amount of5-azacytidine of at least about 300 mg; have an amount of 5-azacytidineof at least about 350 mg; have an amount of 5-azacytidine of at leastabout 400 mg; have an amount of 5-azacytidine of at least about 450 mg;have an amount of 5-azacytidine of at least about 500 mg; have an amountof 5-azacytidine of at least about 600 mg; have an amount of5-azacytidine of at least about 1000 mg; have an amount of 5-azacytidineof about 40 mg; have an amount of 5-azacytidine of about 50 mg; have anamount of 5-azacytidine of about 60 mg; have an amount of 5-azacytidineof about 80 mg; have an amount of 5-azacytidine of about 100 mg; have anamount of 5-azacytidine of about 120 mg; have an amount of 5-azacytidineof about 150 mg; have an amount of 5-azacytidine of about 200 mg; havean amount of 5-azacytidine of about 250 mg; have an amount of5-azacytidine of about 300 mg; have an amount of 5-azacytidine of about350 mg; have an amount of 5-azacytidine of about 400 mg; have an amountof 5-azacytidine of about 450 mg; have an amount of 5-azacytidine ofabout 500 mg; have an amount of 5-azacytidine of about 600 mg; have anamount of 5-azacytidine of about 1000 mg; achieve anarea-under-the-curve value of at least about 200 ng-hr/mL following oraladministration to a subject; achieve an area-under-the-curve value of atleast about 400 ng-hr/mL following oral administration to a subject;achieve a maximum plasma concentration of at least about 100 ng/mLfollowing oral administration to a subject; achieve a maximum plasmaconcentration of at least about 200 ng/mL following oral administrationto a subject; achieve a time to maximum plasma concentration of lessthan about 90 minutes following oral administration to a subject; and/orachieve a time to maximum plasma concentration of less than about 60minutes following oral administration to a subject.

Specific embodiments herein provide, inter alia, methods for treating asubject having cancer or a disease associated with abnormal cellproliferation, comprising orally administering to the subject apharmaceutical composition comprising a therapeutically effective amountof 5-azacytidine. Specific embodiments herein provide, inter alia,methods for treating a subject having cancer or a disease associatedwith abnormal cell proliferation, comprising orally administering to thesubject a pharmaceutical composition comprising a therapeuticallyeffective amount of 5-azacytidine, wherein the composition releases the5-azacytidine substantially in the stomach following oral administrationto the subject. Further embodiments herein provide the aforementionedmethods, in which: the disease is myelodysplastic syndrome; the diseaseis acute myelogenous leukemia; the disease is cancer; the disease is asolid tumor; the disease is a cancer of the bladder, ovary, pancreas,lung, colon, head and neck, breast, or skin; the disease is a cancer ofthe bladder, ovary, pancreas, lung, or colon; the disease is a relapsedor refractory solid tumor; the method further comprises co-administeringto the subject in need thereof an additional therapeutic agent selectedfrom any additional therapeutic agent disclosed herein; the compositionis an immediate release composition; the composition does not have anenteric coating; the composition further comprises a permeationenhancer; the composition further comprises the permeation enhancerd-alpha-tocopheryl polyethylene glycol 1000 succinate; the compositionfurther comprises d-alpha-tocopheryl polyethylene glycol 1000 succinatein the formulation at about 2% by weight relative to the total weight ofthe formulation; the method further comprises not co-administering acytidine deaminase inhibitor with the cytidine analog; the compositionis a single unit dosage form; the composition is a tablet; thecomposition is a capsule; the composition further comprises an excipientselected from any excipient disclosed herein; the amount of5-azacytidine is at least about 40 mg; the amount of 5-azacytidine is atleast about 50 mg; the amount of 5-azacytidine is at least about 60 mg;the amount of 5-azacytidine is at least about 80 mg; the amount of5-azacytidine is at least about 100 mg; the amount of 5-azacytidine isat least about 120 mg; the amount of 5-azacytidine is at least about 150mg; the amount of 5-azacytidine is at least about 200 mg; the amount of5-azacytidine is at least about 250 mg; the amount of 5-azacytidine isat least about 300 mg; the amount of 5-azacytidine is at least about 350mg; the amount of 5-azacytidine is at least about 400 mg; the amount of5-azacytidine is at least about 450 mg; the amount of 5-azacytidine isat least about 500 mg; the amount of 5-azacytidine is at least about 600mg; the amount of 5-azacytidine is at least about 1000 mg; the amount of5-azacytidine is about 40 mg; the amount of 5-azacytidine is about 50mg; the amount of 5-azacytidine is about 60 mg; the amount of5-azacytidine is about 80 mg; the amount of 5-azacytidine is about 100mg; the amount of 5-azacytidine is about 120 mg; the amount of5-azacytidine is about 150 mg; the amount of 5-azacytidine is about 200mg; the amount of 5-azacytidine is about 250 mg; the amount of5-azacytidine is about 300 mg; the amount of 5-azacytidine is about 350mg; the amount of 5-azacytidine is about 400 mg; the amount of5-azacytidine is about 450 mg; the amount of 5-azacytidine is about 500mg; the amount of 5-azacytidine is about 600 mg; the amount of5-azacytidine is about 1000 mg; the method achieves anarea-under-the-curve value of at least about 200 ng-hr/mL following oraladministration to the subject; the method achieves anarea-under-the-curve value of at least about 400 ng-hr/mL following oraladministration to the subject; the method achieves a maximum plasmaconcentration of at least about 100 ng/mL following oral administrationto the subject; the method achieves a maximum plasma concentration of atleast about 200 ng/mL following oral administration to the subject; themethod achieves a time to maximum plasma concentration of less thanabout 90 minutes following oral administration to the subject; and/orthe method achieves a time to maximum plasma concentration of less thanabout 60 minutes following oral administration to the subject.

IV. BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: Dosing and sampling schema for Part 1, Arm A of a clinical studyon orally dosed 5-azacytidine.

FIG. 2: Dosing and sampling schema for Part 1, Arm B of a clinical studyon orally dosed 5-azacytidine.

FIG. 3: Dosing and sampling schema for Part 1, Arm C of a clinical studyon orally dosed 5-azacytidine.

FIG. 4: Dose levels and dose escalation rules for Arms A and B of aclinical study on orally dosed 5-azacytidine.

FIG. 5: Dose levels and dose escalation rules for Arm C of a clinicalstudy on orally dosed 5-azacytidine.

FIG. 6: Modeling of clinical dosing schema in cancer cells.

FIG. 7: Baseline and Cycle 2 CT cross section scans of patients in5-azacytidine and Abraxane treatment group with a pancreatic tumor, andendometrial tumor.

FIG. 8: Baseline and Cycle 2 CT cross section scans of a patient in5-azacytidine treatment group with a nasopharyngeal carcinoma.

FIG. 9: A graph of change in size of nasopharyngeal carcinoma targetlesions in subjects undergoing 5-azacytidine single agent treatment.

V. DETAILED DESCRIPTION

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art. All publications and patents referred to herein areincorporated by reference herein in their entireties.

A. DEFINITIONS

As used in the specification and the accompanying claims, the indefinitearticles “a” and “an” and the definite article “the” include plural aswell as singular referents, unless the context clearly dictatesotherwise.

The term “about” or “approximately” means an acceptable error for aparticular value as determined by one of ordinary skill in the art,which depends in part on how the value is measured or determined. Incertain embodiments, the term “about” or “approximately” means within 1,2, 3, or 4 standard deviations. In certain embodiments, the term “about”or “approximately” means within 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%,5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, or 0.05% of a given value or range.

As used herein, and unless otherwise specified, the terms “treat,”“treating” and “treatment” refer to the eradication or amelioration of adisease or disorder, or of one or more symptoms associated with thedisease or disorder. In certain embodiments, the terms refer tominimizing the spread or worsening of the disease or disorder resultingfrom the administration of one or more prophylactic or therapeuticagents to a subject with such a disease or disorder. In someembodiments, the terms refer to the administration of a compound ordosage form provided herein, with or without one or more additionalactive agent(s), after the onset of symptoms of the particular disease.

As used herein, and unless otherwise specified, the terms “prevent,”“preventing” and “prevention” refer to the prevention of the onset,recurrence or spread of a disease or disorder, or of one or moresymptoms thereof. In certain embodiments, the terms refer to thetreatment with or administration of a compound or dosage form providedherein, with or without one or more other additional active agent(s),prior to the onset of symptoms, particularly to subjects at risk ofdisease or disorders provided herein. The terms encompass the inhibitionor reduction of a symptom of the particular disease. Subjects withfamilial history of a disease are potential candidates for preventiveregimens in certain embodiments. In addition, subjects who have ahistory of recurring symptoms are also potential candidates forprevention. In this regard, the term “prevention” may be interchangeablyused with the term “prophylactic treatment.”

As used herein, and unless otherwise specified, the terms “manage,”“managing” and “management” refer to preventing or slowing theprogression, spread or worsening of a disease or disorder, or of one ormore symptoms thereof. Often, the beneficial effects that a subjectderives from a prophylactic and/or therapeutic agent do not result in acure of the disease or disorder. In this regard, the term “managing”encompasses treating a subject who had suffered from the particulardisease in an attempt to prevent or minimize the recurrence of thedisease.

As used herein, “amelioration” of the symptoms of a particular disorderby administration of a particular pharmaceutical composition refers toany lessening, whether permanent or temporary, lasting or transient,that can be attributed to or associated with administration of thecomposition.

As used herein, and unless otherwise specified, the terms“therapeutically effective amount” and “effective amount” of a compoundmean an amount sufficient to provide a therapeutic benefit in thetreatment or management of a disease or disorder, or to delay orminimize one or more symptoms associated with the disease or disorder. A“therapeutically effective amount” and “effective amount” of a compoundmean an amount of therapeutic agent, alone or in combination with one ormore other agent(s), which provides a therapeutic benefit in thetreatment or management of the disease or disorder. The terms“therapeutically effective amount” and “effective amount” can encompassan amount that improves overall therapy, reduces or avoids symptoms orcauses of disease or disorder, or enhances the therapeutic efficacy ofanother therapeutic agent.

As used herein, and unless otherwise specified, a “prophylacticallyeffective amount” of a compound is an amount sufficient to prevent adisease or disorder, or prevent its recurrence. A prophylacticallyeffective amount of a compound means an amount of therapeutic agent,alone or in combination with one or more other agent(s), which providesa prophylactic benefit in the prevention of the disease. The term“prophylactically effective amount” can encompass an amount thatimproves overall prophylaxis or enhances the prophylactic efficacy ofanother prophylactic agent.

As used herein, and unless otherwise specified, the term “subject” isdefined herein to include animals such as mammals, including, but notlimited to, primates (e.g., humans), cows, sheep, goats, horses, dogs,cats, rabbits, rats, mice, and the like. In specific embodiments, thesubject is a human. The terms “subject” and “patient” are usedinterchangeably herein in reference, for example, to a mammaliansubject, such as a human.

“Tumor,” as used herein, refers to all neoplastic cell growth andproliferation, whether malignant or benign, and all pre-cancerous andcancerous cells and tissues. “Neoplastic,” as used herein, refers to anyform of dysregulated or unregulated cell growth, whether malignant orbenign, resulting in abnormal tissue growth. Thus, “neoplastic cells”include malignant and benign cells having dysregulated or unregulatedcell growth.

As used herein, the terms “cancer” and “cancerous” refer to or describethe physiological condition in mammals that is typically characterizedby unregulated cell growth. Examples of cancer include, but are notlimited to blood-borne (e.g., lymphoma, leukemia) and solid tumors.

As used herein, and unless otherwise specified, the term “proliferative”disorder or disease refers to unwanted cell proliferation of one or moresubset of cells in a multicellular organism resulting in harm (i.e.,discomfort or decreased life expectancy) to the multicellular organism.For example, as used herein, proliferative disorder or disease includesneoplastic disorders and other proliferative disorders.

As used herein, and unless otherwise specified, the term “relapsed”refers to a situation where a subject, that has had a remission ofcancer after a therapy, has a return of cancer cells.

As used herein, and unless otherwise specified, the term “refractory” or“resistant” refers to a circumstance where a subject, even afterintensive treatment, has residual cancer cells in the body.

As used herein, and unless otherwise specified, the term “drugresistance” refers to the condition when a disease does not respond tothe treatment of a drug or drugs. Drug resistance can be eitherintrinsic, which means the disease has never been responsive to the drugor drugs, or it can be acquired, which means the disease ceasesresponding to a drug or drugs that the disease had previously respondedto. In certain embodiments, drug resistance is intrinsic. In certainembodiments, the drug resistance is acquired.

As used herein, and unless otherwise specified, the term “anti-canceragent,” “anticancer agent” or “cancer therapeutic agent” is meant toinclude anti-proliferative agents and chemotherapeutic agents,including, but not limited to, antimetabolites (e.g., 5-fluoro uracil,methotrexate, azacitidine, decitabine, fludarabine, cytarabine (alsoknown as cytosine arabinoside or Ara-C), and high dose cytarabine),antimicrotubule agents (e.g., vinca alkaloids, such as vincristine andvinblastine; and taxanes, such as paclitaxel and docetaxel), alkylatingagents (e.g., mechlorethamine, chlorambucil, cyclophosphamide,melphalan, ifosfamide, carmustine, busulfan, cyclophosphamide,dacarbazine, ifosfamide, and nitrosoureas, such as carmustine,lomustine, bischloroethylnitrosurea, and hydroxyurea), platinum agents(e.g., cisplatin, carboplatin, oxaliplatin, satraplatin (JM-216), andCI-973), anthracyclines (e.g., doxorubicin and daunorubicin), antitumorantibiotics (e.g., mitomycin, bleomycin, idarubicin, adriamycin,daunomycin (also known as daunorubicin, rubidomycin, or cerubidine), andmitoxantrone), topoisomerase inhibitors (e.g., etoposide andcamptothecins), purine antagonists or pyrimidine antagonists (e.g.,6-mercaptopurine, 5-fluorouracil, cytarabine, clofarabine, andgemcitabine), cell maturing agents (e.g., arsenic trioxide andtretinoin), DNA repair enzyme inhibitors (e.g., podophyllotoxines,etoposide, irinotecan, topotecan, and teniposide), enzymes that preventcell survival (e.g., asparaginase and pegaspargase), histone deacetylaseinhibitors (e.g., vorinostat), any other cytotoxic agents (e.g.,estramustine phosphate, dexamethasone, prednimustine, and procarbazine),hormones (e.g., dexamethasone, prednisone, methylprednisolone,tamoxifen, leuprolide, flutamide, and megestrol), monoclonal antibodies(e.g., gemtuzumab ozogamicin, alemtuzumab, rituximab, andyttrium-90-ibritumomab tiuxetan), immuno-modulators (e.g., thalidomideand lenalidomide), Bcr-Abl kinase inhibitors (e.g., AP23464, AZD0530,CGP76030, PD180970, SKI-606, imatinib, BMS354825 (dasatinib), AMN107(nilotinib), and VX-680), hormone agonists or antagonists, partialagonists or partial antagonists, kinase inhibitors, surgery,radiotherapy (e.g., gamma-radiation, neutron bean radiotherapy, electronbeam radiotherapy, proton therapy, brachytherapy, and systemicradioactive isotopes), endocrine therapy, biological response modifiers(e.g., interferons, interleukins, and tumor necrosis factor),hyperthermia and cryotherapy, and agents to attenuate any adverseeffects (e.g., antiemetics).

As used herein, and unless otherwise specified, the terms“co-administration” and “in combination with” include the administrationof two or more therapeutic agents simultaneously, concurrently, orsequentially within no specific time limits unless otherwise indicated.In one embodiment, the agents are present in the cell or in thesubject's body at the same time or exert their biological or therapeuticeffect at the same time. In one embodiment, the therapeutic agents arein the same composition or unit dosage form. In other embodiments, thetherapeutic agents are in separate compositions or unit dosage forms. Incertain embodiments, a first agent can be administered prior to (e.g., 5minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before),essentially concomitantly with, or subsequent to (e.g., 5 minutes, 15minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks,4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) theadministration of a second therapeutic agent.

The terms “composition,” “formulation,” and “dosage form,” as usedherein are intended to encompass compositions comprising the specifiedingredient(s) (in the specified amounts, if indicated), as well as anyproduct(s) which result, directly or indirectly, from combination of thespecified ingredient(s) in the specified amount(s). By “pharmaceutical”or “pharmaceutically acceptable” it is meant that any diluent(s),excipient(s) or carrier(s) in the composition, formulation, or dosageform are compatible with the other ingredient(s) and not deleterious tothe recipient thereof. Unless indicated otherwise, the terms“composition,” “formulation,” and “dosage form” are used hereininterchangeably.

The term “immediate release,” when used herein in reference to acomposition, formulation, or dosage form provided herein, means that thecomposition, formulation, or dosage form does not comprise a component(e.g., a coating) that serves to delay the spatial and/or temporalrelease of some or all of the API from the composition, formulation, ordosage form following oral administration. In certain embodiments, animmediate release composition, formulation, or dosage form is one thatreleases the API substantially in the stomach following oraladministration. In certain embodiments, an immediate releasecomposition, formulation, or dosage form is one that releases the APIsubstantially in the stomach or the upper intestine following oraladministration. In specific embodiments, an immediate releasecomposition, formulation, or dosage form is one that is notdelayed-release. In specific embodiments, an immediate releasecomposition, formulation, or dosage form is one that does not comprisean enteric coating.

The term “non-enteric-coated,” when used herein, refers to apharmaceutical composition, formulation, or dosage form that does notcomprise a coating intended to release the active ingredient(s) beyondthe stomach (e.g., in the intestine). In certain embodiments, anon-enteric-coated composition, formulation, or dosage form is designedto release the active ingredient(s) substantially in the stomach. Incertain embodiments, a non-enteric-coated composition, formulation, ordosage form is designed to release the active ingredient(s)substantially in the stomach and the upper intestine.

The term “substantially in the stomach,” when used herein in referenceto a composition, formulation, or dosage form provided herein, meansthat at least about 99%, at least about 95%, at least about 90%, atleast about 85%, at least about 80%, at least about 75%, at least about70%, at least about 65%, at least about 60%, at least about 55%, atleast about 50%, at least about 45%, at least about 40%, at least about35%, at least about 30%, at least about 25%, at least about 20%, atleast about 15%, or at least about 10% of the cytidine analog isreleased in the stomach. The term “released in the stomach” and relatedterms as used herein refer to the process whereby the cytidine analog ismade available for uptake by or transport across cells lining thestomach and then made available to the body.

The term “isotopic composition” refers to the amount of each isotopepresent in a given atomic position, and “natural isotopic composition”refers to the naturally occurring isotopic composition or abundance fora given atomic position. Atomic positions containing their naturalisotopic composition may also be referred to herein as “non-enriched.”Unless otherwise designated, the atomic positions of the compoundsrecited herein are meant to represent any stable isotope of that atom.For example, unless otherwise stated, when a position is designatedspecifically as “H” or “hydrogen,” the position is understood to havehydrogen at its natural isotopic composition.

The term “isotopically enriched” refers to an atomic position having anisotopic composition other than the natural isotopic composition of thatatom. “Isotopically enriched” may also refer to a compound containing atleast one atomic position having an isotopic composition other than thenatural isotopic composition of that atom. As used herein, an“isotopologue” is an isotopically enriched compound.

The term “isotopic enrichment” refers to the percentage of incorporationof an amount of a specific isotope at a given atomic position in amolecule in the place of that atom's natural isotopic composition. Forexample, deuterium enrichment of 1% at a given position means that 1% ofthe molecules in a given sample contain deuterium at the specifiedposition. Because the naturally occurring distribution of deuterium isabout 0.0156%, deuterium enrichment at any position in a compoundsynthesized using non-enriched starting materials is about 0.0156%.

The term “isotopic enrichment factor” refers to the ratio between theisotopic composition and the natural isotopic composition of a specifiedisotope.

With regard to the compounds provided herein, when a particular atomicposition is designated as having deuterium or “D,” it is understood thatthe abundance of deuterium at that position is substantially greaterthan the natural abundance of deuterium, which is about 0.015%. Aposition designated as having deuterium typically has a minimum isotopicenrichment factor of, in particular embodiments, at least 1000 (15%deuterium incorporation), at least 2000 (30% deuterium incorporation),at least 3000 (45% deuterium incorporation), at least 3500 (52.5%deuterium incorporation), at least 4000 (60% deuterium incorporation),at least 4500 (67.5% deuterium incorporation), at least 5000 (75%deuterium incorporation), at least 5500 (82.5% deuterium incorporation),at least 6000 (90% deuterium incorporation), at least 6333.3 (95%deuterium incorporation), at least 6466.7 (97% deuterium incorporation),at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5%deuterium incorporation) at each designated deuterium position.

The isotopic enrichment and isotopic enrichment factor of the compoundsprovided herein can be determined using conventional analytical methodsknown to one of ordinary skill in the art, including, e.g., massspectrometry, nuclear magnetic resonance spectroscopy, andcrystallography.

As used herein, and unless otherwise specified, the term“pharmaceutically acceptable carrier,” “pharmaceutically acceptableexcipient,” “physiologically acceptable carrier,” or “physiologicallyacceptable excipient” refers to a pharmaceutically-acceptable material,composition, or vehicle, such as, e.g., a liquid or solid filler,diluent, excipient, solvent, or encapsulating material. In oneembodiment, each component is “pharmaceutically acceptable” in the senseof being compatible with the other ingredients of a pharmaceuticalformulation, and suitable for use in contact with the tissue or organ ofhumans and animals without excessive toxicity, irritation, allergicresponse, immunogenicity, or other problems or complications,commensurate with a reasonable benefit/risk ratio. In one embodiment, by“pharmaceutical” or “pharmaceutically acceptable” it is meant that anydiluent(s), excipient(s) or carrier(s) in the composition, formulation,or dosage form are compatible with the other ingredient(s) and notdeleterious to the recipient thereof. See, e.g., Remington, The Scienceand Practice of Pharmacy, 21st Edition; Lippincott Williams & Wilkins:Philadelphia, Pa., 2005; Handbook of Pharmaceutical Excipients, 5thEdition; Rowe et al., ed., The Pharmaceutical Press and the AmericanPharmaceutical Association: 2005; and Handbook of PharmaceuticalAdditives, 3rd Edition; Ash and Ash ed., Gower Publishing Company: 2007;Pharmaceutical Preformulation and Formulation, Gibson ed., CRC PressLLC: Boca Raton, Fla., 2004.

As used herein, and unless otherwise specified, the term “hydrate” meansa compound provided herein or a salt thereof, which further includes astoichiometric or non-stoichiometric amount of water bound bynon-covalent intermolecular forces.

As used herein, and unless otherwise specified, the term “solvate” meansa solvate formed from the association of one or more solvent moleculesto a compound provided herein. The term “solvate” includes hydrates(e.g., mono-hydrate, dihydrate, trihydrate, tetrahydrate and the like).

As used herein, and unless otherwise specified, a compound describedherein is intended to encompass all possible stereoisomers, unless aparticular stereochemistry is specified. Where structural isomers of acompound are interconvertible via a low energy barrier, the compound mayexist as a single tautomer or a mixture of tautomers. This can take theform of proton tautomerism; or so-called valence tautomerism in thecompound, e.g., that contain an aromatic moiety.

B. CYTIDINE ANALOGS

1. Overview

Provided herein are dosage forms, pharmaceutical formulations, andcompositions comprising cytidine analogs that release the APIsubstantially in the stomach upon oral administration. In certainembodiments, the cytidine analog is 5-azacytidine. In certainembodiments, the cytidine analog is 5-aza-2′-deoxycytidine (decitabineor 5-aza-CdR). In certain embodiments, the cytidine analog is, forexample: 1-β-D-arabinofuranosylcytosine (Cytarabine or ara-C);pseudoiso-cytidine (psi ICR); 5-fluoro-2′-deoxycytidine (FCdR);2′-deoxy-2′,2′-difluorocytidine (Gemcitabine);5-aza-2′-deoxy-2′,2′-difluorocytidine; 5-aza-2′-deoxy-2′-fluorocytidine;1-β-D-ribofuranosyl-2(1H)-pyrimidinone (Zebularine);2′,3′-dideoxy-5-fluoro-3′-thiacytidine (Emtriva); 2′-cyclocytidine(Ancitabine); 1-β-D-arabinofuranosyl-5-azacytosine (Fazarabine orara-AC); 6-azacytidine (6-aza-CR); 5,6-dihydro-5-azacytidine(dH-aza-CR); N⁴-pentyloxy-carbonyl-5′-deoxy-5-fluorocytidine(Capecitabine); N⁴-octadecyl-cytarabine; elaidic acid cytarabine; or aconjugated compound comprising a cytidine analog and a fatty acid (e.g.,an azacitidine—fatty acid conjugate, including, but not limited to,CP-4200 (Clavis Pharma ASA) or a compound disclosed in WO 2009/042767,such as aza-C-5′-petroselinic acid ester or aza-C-5′-petroselaidic acidester).

In certain embodiments, cytidine analogs provided herein includeesterified derivatives of cytidine analogs, such as, e.g., esterifiedderivatives of 5-azacytidine. In particular embodiments, esterifiedderivatives are cytidine analogs that contain an ester moiety (e.g., anacetyl group) at one or more positions on the cytidine analog molecule.Esterified derivatives may be prepared by any method known in the art.In certain embodiments, esterified derivatives of a cytidine analogserve as prodrugs of the cytidine analog, such that, e.g., followingadministration of an esterified derivative, the derivative isdeacetylated in vivo to yield the cytidine analog. A particularembodiment herein provides 2′,3′,5′-triacetyl-5-azacytidine (TAC), whichpossesses favorable physical-chemical and therapeutic properties. See,e.g., International Publication No. WO 2008/092127 (InternationalApplication No. PCT/US2008/052124); Ziemba, A. J., et al., “Developmentof Oral Demethylating Agents for the Treatment of MyelodysplasticSyndrome” (Abstract No. 3369), In: Proceedings of the 100th AnnualMeeting of the American Association for Cancer Research; 2009 Apr.18-22; Denver, Co. Philadelphia (Pa.): AACR; 2009 (both of which areincorporated by reference herein in their entireties).

In certain embodiments, the cytidine analogs provided herein include anycompound which is structurally related to cytidine or deoxycytidine andfunctionally mimics and/or antagonizes the action of cytidine ordeoxycytidine. Certain embodiments herein provide salts, cocrystals,solvates (e.g., hydrates), complexes, prodrugs, precursors, metabolites,and/or other derivatives of the cytidine analogs provided herein. Forexample, particular embodiments provide salts, cocrystals, solvates(e.g., hydrates), complexes, precursors, metabolites, and/or otherderivatives of 5-azacytidine. Certain embodiments provide cytidineanalogs that are not salts, cocrystals, solvates (e.g., hydrates), orcomplexes of the cytidine analogs provided herein. For example,particular embodiments provide 5-azacytidine in a non-ionized,non-solvated (e.g., anhydrous), non-complexed form. Certain embodimentsherein provide mixtures of two or more cytidine analogs provided herein.

Cytidine analogs provided herein may be prepared using synthetic methodsand procedures referenced herein or otherwise available in theliterature. For example, particular methods for synthesizing5-azacytidine are taught in, e.g., U.S. Pat. No. 7,038,038 andreferences discussed therein, each of which is incorporated herein byreference. 5-Azacytidine is also available from Celgene Corporation,Warren, N.J. Other cytidine analogs provided herein may be preparedusing previously disclosed synthetic procedures available to a person ofordinary skill in the art.

It should be noted that if there is a discrepancy between a depictedstructure and a chemical name given that structure, the depictedstructure is to be accorded more weight. In addition, if thestereochemistry of a structure or a portion of a structure is notindicated with, for example, bold or dashed lines, the structure orportion of the structure is to be interpreted as encompassing allstereoisomers. Where the compound provided herein contains an alkenyl oralkenylene group, the compound may exist as one geometric (i.e.,cis/trans or E/Z) isomer or a mixture of geometric (i.e., cis/trans orE/Z) isomers. Unless otherwise specified, a compound provided herein isintended to encompass all geometric isomers.

Where structural isomers are inter-convertible, the compound may existas a single tautomer or a mixture of tautomers. This can take the formof proton tautomerism in the compound that contains, for example, animino, keto, or oxime group; or so-called valence tautomerism in thecompound that contain, for example, an aromatic moiety. It follows thata single compound may exhibit more than one type of isomerism. It willbe understood that unless otherwise specified, a compound providedherein is intended to encompass all possible tautomers. Similarly,unless otherwise specified, a compound provided herein is intended toencompass all possible stereoisomers.

The compounds provided herein may be enantiomerically pure, such as asingle enantiomer or a single diastereomer, or be stereoisomericmixtures, such as a mixture of enantiomers, e.g., a racemic mixture oftwo enantiomers; or a mixture of two or more diastereomers. Conventionaltechniques for the preparation/isolation of individual enantiomersinclude synthesis from a suitable optically pure precursor, asymmetricsynthesis from achiral starting materials, or resolution of anenantiomeric mixture, for example, by chiral chromatography,recrystallization, resolution, diastereomeric salt formation, orderivatization into diastereomeric adducts followed by separation. Insome instances, for compounds that undergo epimerization in vivo, one ofskill in the art will recognize that administration of a compound in its(R) form is equivalent to administration of the compound in its (S)form, and vice versa.

When the compound provided herein contains an acidic or basic moiety, itmay also be provided as a pharmaceutically acceptable salt (See, Bergeet al., J. Pharm. Sci. 1977, 66, 1-19; and “Handbook of PharmaceuticalSalts, Properties, and Use,” Stahl and Wermuth, Ed.; Wiley-VCH and VHCA,Zurich, 2002).

Suitable acids for use in the preparation of pharmaceutically acceptablesalts include, but are not limited to, acetic acid, 2,2-dichloroaceticacid, acylated amino acids, adipic acid, alginic acid, ascorbic acid,L-aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoicacid, boric acid, (+)-camphoric acid, camphorsulfonic acid,(+)-(1S)-camphor-10-sulfonic acid, capric acid, caproic acid, caprylicacid, cinnamic acid, citric acid, cyclamic acid, cyclohexanesulfamicacid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonicacid, 2-hydroxy-ethanesulfonic acid, formic acid, fumaric acid,galactaric acid, gentisic acid, glucoheptonic acid, D-gluconic acid,D-glucuronic acid, L-glutamic acid, α-oxoglutaric acid, glycolic acid,hippuric acid, hydrobromic acid, hydrochloric acid, hydroiodic acid,(+)-L-lactic acid, (±)-DL-lactic acid, lactobionic acid, lauric acid,maleic acid, (−)-L-malic acid, malonic acid, (±)-DL-mandelic acid,methanesulfonic acid, naphthalene-2-sulfonic acid,naphthalene-1,5-disulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinicacid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid,pamoic acid, perchloric acid, phosphoric acid, L-pyroglutamic acid,saccharic acid, salicylic acid, 4-amino-salicylic acid, sebacic acid,stearic acid, succinic acid, sulfuric acid, tannic acid, (+)-L-tartaricacid, thiocyanic acid, p-toluenesulfonic acid, undecylenic acid, andvaleric acid.

Suitable bases for use in the preparation of pharmaceutically acceptablesalts, including, but not limited to, inorganic bases, such as magnesiumhydroxide, calcium hydroxide, potassium hydroxide, zinc hydroxide, orsodium hydroxide; and organic bases, such as primary, secondary,tertiary, and quaternary, aliphatic and aromatic amines, includingL-arginine, benethamine, benzathine, choline, deanol, diethanolamine,diethylamine, dimethylamine, dipropylamine, diisopropylamine,2-(diethylamino)-ethanol, ethanolamine, ethylamine, ethylenediamine,isopropylamine, N-methyl-glucamine, hydrabamine, 1H-imidazole, L-lysine,morpholine, 4-(2-hydroxyethyl)-morpholine, methylamine, piperidine,piperazine, propylamine, pyrrolidine, 1-(2-hydroxyethyl)-pyrrolidine,pyridine, quinuclidine, quinoline, isoquinoline, secondary amines,triethanolamine, trimethylamine, triethylamine, N-methyl-D-glucamine,2-amino-2-(hydroxymethyl)-1,3-propanediol, and tromethamine.

The compound provided herein may also be provided as a prodrug, which isa functional derivative of a compound provided herein, and is readilyconvertible into the parent compound in vivo. Prodrugs are often usefulbecause, in some situations, they may be easier to administer than theparent compound. They may, for instance, be bioavailable by oraladministration whereas the parent compound is not. The prodrug may alsohave enhanced solubility in pharmaceutical compositions over the parentcompound. A prodrug may be converted into the parent drug by variousmechanisms, including enzymatic processes and metabolic hydrolysis. SeeHarper, Progress in Drug Research 1962, 4, 221-294; Morozowich et al. in“Design of Biopharmaceutical Properties through Prodrugs and Analogs,”Roche Ed., APHA Acad. Pharm. Sci. 1977; “Bioreversible Carriers in Drugin Drug Design, Theory and Application,” Roche Ed., APHA Acad. Pharm.Sci. 1987; “Design of Prodrugs,” Bundgaard, Elsevier, 1985; Wang et al.,Curr. Pharm. Design 1999, 5, 265-287; Pauletti et al., Adv. Drug.Delivery Rev. 1997, 27, 235-256; Mizen et al., Pharm. Biotech. 1998, 11,345-365; Gaignault et al., Pract. Med. Chem. 1996, 671-696; Asgharnejadin “Transport Processes in Pharmaceutical Systems,” Amidon et al., Ed.,Marcell Dekker, 185-218, 2000; Balant et al., Eur. J. Drug Metab.Pharmacokinet. 1990, 15, 143-53; Balimane and Sinko, Adv. Drug DeliveryRev. 1999, 39, 183-209; Browne, Clin. Neuropharmacol. 1997, 20, 1-12;Bundgaard, Arch. Pharm. Chem. 1979, 86, 1-39; Bundgaard, Controlled DrugDelivery 1987, 17, 179-96; Bundgaard, Adv. Drug Delivery Rev. 1992, 8,1-38; Fleisher et al., Adv. Drug Delivery Rev. 1996, 19, 115-130;Fleisher et al., Methods Enzymol. 1985, 112, 360-381; Farquhar et al.,J. Pharm. Sci. 1983, 72, 324-325; Freeman et al., J. Chem. Soc., Chem.Commun. 1991, 875-877; Friis and Bundgaard, Eur. J. Pharm. Sci. 1996, 4,49-59; Gangwar et al., Des. Biopharm. Prop. Prodrugs Analogs, 1977,409-421; Nathwani and Wood, Drugs 1993, 45, 866-94; Sinhababu andThakker, Adv. Drug Delivery Rev. 1996, 19, 241-273; Stella et al., Drugs1985, 29, 455-73; Tan et al., Adv. Drug Delivery Rev. 1999, 39, 117-151;Taylor, Adv. Drug Delivery Rev. 1996, 19, 131-148; Valentino andBorchardt, Drug Discovery Today 1997, 2, 148-155; Wiebe and Knaus, Adv.Drug Delivery Rev. 1999, 39, 63-80; and Waller et al., Br. J. Clin.Pharmac. 1989, 28, 497-507.

In certain embodiments, exemplary cytidine analogs have the structuresprovided below:

2. Isotopically Enriched Cytidine Analogs

Particular embodiments herein provide isotopically enriched cytidineanalogs, prodrugs thereof, synthetic intermediates thereof, andmetabolites thereof. For example, specific embodiments herein provideisotopically enriched 5-azacytidine.

Isotopic enrichment (e.g., deuteration) of pharmaceuticals to improvepharmacokinetics (“PK”), pharmacodynamics (“PD”), and toxicity profiles,has been demonstrated previously with some classes of drugs. See, e.g.,Lijinsky et. al., Food Cosmet. Toxicol., 20: 393 (1982); Lijinsky et.al., J. Nat. Cancer Inst., 69: 1127 (1982); Mangold et. al., MutationRes. 308: 33 (1994); Gordon et. al., Drug Metab. Dispos., 15: 589(1987); Zello et. al., Metabolism, 43: 487 (1994); Gately et. al., J.Nucl. Med., 27: 388 (1986); Wade, D., Chem. Biol. Interact. 117: 191(1999).

Without being limited by any particular theory, isotopic enrichment of adrug can be used, for example, to: (1) reduce or eliminate unwantedmetabolites; (2) increase the half-life of the parent drug; (3) decreasethe number of doses needed to achieve a desired effect; (4) decrease theamount of a dose necessary to achieve a desired effect; (5) increase theformation of active metabolites, if any are formed; and/or (6) decreasethe production of deleterious metabolites in specific tissues and/orcreate a more effective drug and/or a safer drug for combinationtherapy, whether the combination therapy is intentional or not.

Replacement of an atom for one of its isotopes may often result in achange in the reaction rate of a chemical reaction. This phenomenon isknown as the Kinetic Isotope Effect (“KIE”). For example, if a C—H bondis broken during a rate-determining step in a chemical reaction (i.e.the step with the highest transition state energy), substitution of adeuterium for that hydrogen will cause a decrease in the reaction rateand the process will slow down. This phenomenon is known as theDeuterium Kinetic Isotope Effect (“DKIE”). See, e.g., Foster et al.,Adv. Drug Res., vol. 14, pp. 1-36 (1985); Kushner et al., Can. J.Physiol. Pharmacol., vol. 77, pp. 79-88 (1999).

Certain embodiments herein provide deuterium enriched 5-azacytidineanalogs, wherein one or more hydrogen(s) in the 5-azacytidine moleculeis/are isotopically enriched with deuterium. In certain embodiments,provided herein are compounds of formula (I):

wherein one or more Y atom(s) (i.e., Y¹, Y², Y³, Y⁴, Y⁵, Y⁶, and Y⁷)is/are hydrogen(s) isotopically enriched with deuterium, and anyremaining Y atom(s) is/are non-enriched hydrogen atom(s). In particularembodiments, one, two, three, four, five, six, or seven of the indicatedY atom(s) is/are isotopically enriched with deuterium, and any remainingY atom(s) is/are non-enriched hydrogen(s).

In certain embodiments, one or more Y atoms on the ribose moiety ofCompound (I) are deuterium-enriched. Particular examples include, butare not limited to, the following compounds, in which the label “D”indicates a deuterium-enriched atomic position, i.e., a samplecomprising the given compound has a deuterium enrichment at theindicated position(s) above the natural abundance of deuterium:

In certain embodiments, the Y atom on the 5-azacytosine moiety ofCompound (I) is deuterium-enriched. Particular example includes thefollowing compound, in which the label “D” indicates adeuterium-enriched atomic position, i.e., a sample comprising the givencompound has a deuterium enrichment at the indicated position(s) abovethe natural abundance of deuterium:

In certain embodiments, one or more Y atoms on the ribose moiety and theY atom on the 5-azacytosine moiety of Compound (I) aredeuterium-enriched. Particular examples include, but are not limited to,the following compounds, in which the label “D” indicates adeuterium-enriched atomic position, i.e., a sample comprising the givencompound has a deuterium enrichment at the indicated position(s) abovethe natural abundance of deuterium:

It is understood that one or more deuterium(s) may exchange withhydrogen under physiological conditions.

Certain embodiments herein provide carbon-13 enriched analogs of5-azacytidine, wherein one or more carbon(s) in the 5-azacytidinemolecule is/are isotopically enriched with carbon-13. In certainembodiments, provided herein are compounds of formula (II):

wherein one or more of 1, 2, 3, 4, 5, 6, 7, or 8 is/are carbon atom(s)isotopically enriched with carbon-13, and any remaining atom(s) of 1, 2,3, 4, 5, 6, 7, or 8 is/are non-enriched carbon atom(s). In particularembodiments, one, two, three, four, five, six, seven, or eight carbonatom(s) (i.e., atoms 1, 2, 3, 4, 5, 6, 7, and 8) is/are isotopicallyenriched with carbon-13, and any remaining carbon atom(s) is/arenon-enriched.

In certain embodiments, one or more carbon atom(s) of the ribose moietyof Compound (II) are enriched with carbon-13. Particular examplesinclude, but are not limited to, the following compounds, in which theasterisk (“*”) indicates a carbon-13 enriched atomic position, i.e., asample comprising the given compound has a carbon-13 enrichment at theindicated position(s) above the natural abundance of carbon-13:

In certain embodiments, one or more carbon atom(s) of the 5-azacytosinemoiety of Compound (II) are enriched with carbon-13. Particular examplesinclude, but are not limited to, the following compounds, in which theasterisk “*” indicates a carbon-13 enriched atomic position, i.e., asample comprising the given compound has a carbon-13 enrichment at theindicated position(s) above the natural abundance of carbon-13:

In certain embodiments, one or more carbon atoms on the ribose moietyand one or more carbon atoms on the 5-azacytosine moiety of Compound(II) are enriched with carbon-13, i.e., any combination of carbon-13enrichment for the ribose moiety and carbon-13 enrichment for theazacitosine moiety is encompassed herein.

In certain embodiments, one or more hydrogen(s) is/are enriched withdeuterium(s) and one or more carbon(s) is/are enriched with carbon-13,i.e., any combination of deuterium enrichment and carbon-13 enrichmentof 5-azacytidine is encompassed herein.

The compounds described herein may be synthesized using any method knownto one of ordinary skill in the art. For example, particular compoundsdescribed herein are synthesized using standard synthetic organicchemistry techniques known to those of ordinary skill in the art. Insome embodiments, known procedures for the synthesis of 5-azacytidineare employed, wherein one or more of the reagents, starting materials,precursors, or intermediates are replaced by one or moreisotopically-enriched reagents, starting materials, precursors, orintermediates, including but not limited to one or moredeuterium-enriched reagents, starting materials, precursors, orintermediates, and/or one or more carbon-13-enriched reagents, startingmaterials, precursors, or intermediates. Isotopically enriched reagents,starting materials, precursors, or intermediates are commerciallyavailable or may be prepared by routine chemical reactions known to oneof skill in the art. In some embodiments, the routes are based on thosedisclosed in U.S. Pat. No. 7,038,038 and U.S. Patent Publication No.2009/0286752 (application Ser. No. 12/466,213), both of which areincorporated herein by reference in their entireties.

C. PHARMACEUTICAL COMPOSITIONS

In one embodiment, provided herein are pharmaceutical compositions,which comprise a cytidine analog, or a pharmaceutically acceptable salt,solvate, or hydrate thereof, as an active ingredient, in combinationwith one or more pharmaceutically acceptable excipient or carrier. Inone embodiment, the pharmaceutical composition comprises at least onenonrelease controlling excipient or carrier. In one embodiment, thepharmaceutical composition comprises at least one release controllingand at least one nonrelease controlling excipient or carrier.

In certain embodiments, the cytidine analog used in the pharmaceuticalcompositions provided herein is in a solid form. Suitable solid formsinclude, but are not limited to, solid forms comprising the free base ofthe cytidine analog, and solid forms comprising salts of the cytidineanalog. In certain embodiments, solid forms provided herein includepolymorphs, solvates (including hydrates), and cocrystals comprising thecytidine analog and/or salts thereof. In certain embodiments, the solidform is a crystal form of the cytidine analog, or a pharmaceuticallyacceptable salt, solvate, or hydrate thereof.

In one embodiment, the pharmaceutical compositions provided herein maybe formulated in various dosage forms for oral, parenteral, and topicaladministration. The pharmaceutical compositions may also be formulatedas modified release dosage forms, including delayed-, extended-,prolonged-, sustained-, pulsed-, controlled-, accelerated- and fast-,targeted-, programmed-release, and gastric retention dosage forms. Thesedosage forms can be prepared according to conventional methods andtechniques known to those skilled in the art. See, e.g., Remington, TheScience and Practice of Pharmacy, 21st Edition; Lippincott Williams &Wilkins: Philadelphia, Pa., 2005; Modified-Release Drug DeliveryTechnology, Rathbone et al., eds., Drugs and the Pharmaceutical Science,Marcel Dekker, Inc.: New York, N.Y., 2003; Vol. 126.

In one embodiment, the pharmaceutical compositions are provided in adosage form for oral administration. In another embodiment, thepharmaceutical compositions are provided in a dosage form for parenteraladministration. In yet another embodiment, the pharmaceuticalcompositions are provided in a dosage form for topical administration.

In one embodiment, the pharmaceutical compositions provided herein maybe provided in a unit-dosage form or multiple-dosage form. A unit-dosageform, as used herein, refers to a physically discrete unit suitable foradministration to human and animal subjects, and packaged individuallyas is known in the art. Each unit-dose contains a predetermined quantityof the active ingredient(s) sufficient to produce the desiredtherapeutic effect, in association with the required pharmaceuticalcarriers or excipients. Examples of a unit-dosage form include anampoule, syringe, and individually packaged tablet and capsule. Aunit-dosage form may be administered in fractions or multiples thereof.A multiple-dosage form is a plurality of identical unit-dosage formspackaged in a single container to be administered in segregatedunit-dosage form. Examples of a multiple-dosage form include a vial,bottle of tablets or capsules, or bottle of pints or gallons.

In one embodiment, the pharmaceutical compositions provided herein maybe administered once or multiple times, at particular intervals of time.It is understood that the precise dosage and duration of treatment mayvary with the age, weight, and condition of the patient being treated,and may be determined empirically using known testing protocols or byextrapolation from in vivo or in vitro test or diagnostic data. It isfurther understood that for any particular individual, specific dosageregimens should be adjusted over time according to the individual needand the professional judgment of the person administering or supervisingthe administration of the formulations.

1. Overview of Oral Dosage Forms

In one embodiment, the pharmaceutical compositions provided herein maybe provided in solid, semisolid, or liquid dosage forms for oraladministration. As used herein, oral administration also includesbuccal, lingual, and sublingual administration. Suitable oral dosageforms include, but are not limited to, tablets, capsules, pills,troches, lozenges, pastilles, cachets, pellets, medicated chewing gum,granules, bulk powders, effervescent or non-effervescent powders orgranules, solutions, emulsions, suspensions, solutions, wafers,sprinkles, elixirs, and syrups. In addition to the active ingredient(s),the pharmaceutical compositions may contain one or more pharmaceuticallyacceptable carriers or excipients, including, but not limited to,binders, fillers, diluents, disintegrants, wetting agents, lubricants,glidants, coloring agents, dye-migration inhibitors, sweetening agents,and flavoring agents.

In one embodiment, provided herein are pharmaceutical formulations andcompositions comprising a cytidine analog (e.g., 5-azacytidine oranother cytidine analog provided herein), and optionally a permeationenhancer, wherein the formulations and compositions are prepared fororal administration. In a particular embodiment, the formulations andcompositions are prepared for release of the cytidine analogsubstantially in the stomach. In specific embodiments, the cytidineanalog (e.g., 5-azacytidine or another cytidine analog provided herein)and the pharmaceutical formulation and composition are used fortreating, preventing, or managing diseases and disorders associated withabnormal cell proliferation, for example, a solid tumor, wherein thecytidine analog, the formulation and composition are prepared for oraladministration, preferably for release of the cytidine analogsubstantially in the stomach. Particular embodiments relate to the useof one or more cytidine analogs (e.g., 5-azacytidine or another cytidineanalog provided herein) for the preparation of pharmaceuticalformulations and compositions for treating particular medicalindications, as provided herein. The pharmaceutical formulations andcompositions comprising a cytidine analog provided herein are intendedfor oral delivery of the cytidine analog in subjects in need thereof.Oral delivery formats include, but are not limited to, tablets,capsules, caplets, solutions, suspensions, and syrups, and may alsocomprise a plurality of granules, beads, powders or pellets that may ormay not be encapsulated. Such formats may also be referred to herein asthe “drug core” which contains the cytidine analog.

Particular embodiments herein provide solid oral dosage forms that aretablets or capsules. In certain embodiments, the formulation is a tabletcomprising a cytidine analog. In certain embodiments, the formulation isa capsule comprising a cytidine analog. In certain embodiments, thetablets or capsules provided herein optionally comprise one or moreexcipients, such as, for example, glidants, diluents, lubricants,colorants, disintegrants, granulating agents, binding agents, polymers,and coating agents. In certain embodiments, the formulation is animmediate release tablet. In certain embodiments, the formulation is acontrolled release tablet releasing the API, e.g., substantially in thestomach. In certain embodiments, the formulation is a hard gelatincapsule. In certain embodiments, the formulation is a soft gelatincapsule. In certain embodiments, the capsule is a hydroxypropylmethylcellulose (HPMC) capsule. In certain embodiments, the formulationis an immediate release capsule. In certain embodiments, the formulationis an immediate or controlled release capsule releasing the API, e.g.,substantially in the stomach. In certain embodiments, the formulation isa rapidly disintegrating tablet that dissolves substantially in themouth following administration. In certain embodiments, embodimentsherein encompass the use of a cytidine analog (e.g., 5-azacytidine oranother cytidine analog provided herein) for the preparation of apharmaceutical composition for treating a disease associated withabnormal cell proliferation, wherein the composition is prepared fororal administration.

2. Performance of Certain Dosage Forms Provided Herein

In certain embodiments, the formulations comprising a cytidine analog,such as, for example, 5-azacytidine or another cytidine analog providedherein, effect an immediate release of the API upon oral administration.In particular embodiments, the formulations comprising a cytidineanalog, such as, for example, 5-azacytidine or another cytidine analogprovided herein, comprise a therapeutically or prophylacticallyeffective amount of the cytidine analog (and, optionally, one or moreexcipients) and effect an immediate release of the API upon oraladministration.

In certain embodiments, the formulations comprising a cytidine analog,such as, for example, 5-azacytidine or another cytidine analog providedherein, effect a controlled release of the API substantially in thestomach upon oral administration. In certain embodiments, theformulations comprising a cytidine analog, such as, for example,5-azacytidine or another cytidine analog provided herein, comprise atherapeutically or prophylactically effective amount of the cytidineanalog and a drug release controlling component which is capable ofreleasing the cytidine analog substantially in the stomach. In certainembodiments, matrices (e.g., polymer matrices) may be employed in theformulation to control the release of the cytidine analog. In certainembodiments, coatings and/or shells may be employed in the formulationto control the release of the cytidine analog in the substantially inthe stomach.

In certain embodiments, the formulations comprising a cytidine analog,such as, for example, 5-azacytidine or another cytidine analog providedherein, release the API substantially in the stomach upon oraladministration. In certain embodiments, the formulations effect animmediate release of the cytidine analog upon oral administration. Incertain embodiments, the formulations optionally further comprises adrug release controlling component, wherein the drug release controllingcomponent is adjusted such that the release of the cytidine analogoccurs substantially in the stomach. In particular embodiments, the drugrelease controlling component is adjusted such that the release of thecytidine analog is immediate and occurs substantially in the stomach. Inparticular embodiments, the drug release controlling component isadjusted such that the release of the cytidine analog is sustained andoccurs substantially in the stomach. In certain embodiments, theformulation of a cytidine analog, such as, for example, 5-azacytidine oranother cytidine analog provided herein, releases the API substantiallyin the stomach, and, subsequently, releases the remainder of the API inthe intestine upon oral administration.

Methods by which skilled practitioners can assess where a drug isreleased in the gastrointestinal tract of a subject are known in theart, and include, for example, scintigraphic studies, testing in abio-relevant medium which simulates the fluid in relevant portions ofthe gastrointestinal tract, among other methods.

Particular embodiments herein provide pharmaceutical formulations (e.g.,immediate release oral formulations and/or formulations that release theAPI substantially in the stomach) comprising a cytidine analog (e.g.,5-azacytidine or another cytidine analog provided herein) that achieve aparticular exposure in the subject to which the formulation is orallyadministered, as compared to a SC dose of the same cytidine analog.Particular embodiments provide oral formulations that achieve anexposure of at least about 5%, at least about 10%, at least about 15%,at least about 20%, at least about 25%, at least about 30%, at leastabout 35%, at least about 40%, at least about 45%, at least about 50%,at least about 55%, at least about 60%, at least about 65%, at leastabout 70%, at least about 75%, at least about 80%, at least about 85%,at least about 90%, at least about 95%, or about 100%, as compared to aSC dose.

In certain embodiments, the formulation (e.g., immediate release oralformulation and/or formulation that release the API substantially in thestomach) comprising a cytidine analog, such as, for example,5-azacytidine or another cytidine analog provided herein, renders acertain percentage of the cytidine analog in the formulationsystemically bioavailable upon oral administration. In certainembodiments, after the subject is orally administered the formulation,the cytidine analog in the formulation is absorbed substantially in thestomach, and becomes available to the body through systemic exposure. Inparticular embodiments, the oral bioavailability of a formulationcomprising a cytidine analog provided herein is, e.g., greater thanabout 1%, greater than about 5%, greater than about 10%, greater thanabout 15%, greater than about 20%, greater than about 25%, greater thanabout 30%, greater than about 35%, greater than about 40%, greater thanabout 45%, greater than about 50%, greater than about 55%, greater thanabout 60%, greater than about 65%, greater than about 70%, greater thanabout 75%, greater than about 80%, greater than about 85%, greater thanabout 90%, greater than about 95%, or about 100%, of the total amount ofthe cytidine analog in the formulation.

Methods by which skilled practitioners can assess the oralbioavailability of a drug formulation in a subject are known in the art.Such methods, include, for example, comparing certain dosing-relatedparameters, such as, but not limited to, maximum plasma concentration(“Cmax”), time to maximum plasma concentration (“Tmax”), orarea-under-the-curve (“AUC”) determinations.

Particular embodiments herein provide pharmaceutical formulations (e.g.,immediate release oral formulations and/or formulations that release theAPI substantially in the stomach) comprising a cytidine analog (e.g.,5-azacytidine or another cytidine analog provided herein) that achieve aparticular AUC value (e.g., AUC(0-t) or AUC(0-∞)) in the subject (e.g.,human) to which the formulation is orally administered. Particularembodiments provide oral formulations that achieve an AUC value of atleast about 25 ng-hr/mL, at least about 50 ng-hr/mL, at least about 75ng-hr/mL, at least about 100 ng-hr/mL, at least about 150 ng-hr/mL, atleast about 200 ng-hr/mL, at least about 250 ng-hr/mL, at least about300 ng-hr/mL, at least about 350 ng-hr/mL, at least about 400 ng-hr/mL,at least about 450 ng-hr/mL, at least about 500 ng-hr/mL, at least about550 ng-hr/mL, at least about 600 ng-hr/mL, at least about 650 ng-hr/mL,at least about 700 ng-hr/mL, at least about 750 ng-hr/mL, at least about800 ng-hr/mL, at least about 850 ng-hr/mL, at least about 900 ng-hr/mL,at least about 950 ng-hr/mL, at least about 1000 ng-hr/mL, at leastabout 1100 ng-hr/mL, at least about 1200 ng-hr/mL, at least about 1300ng-hr/mL, at least about 1400 ng-hr/mL, at least about 1500 ng-hr/mL, atleast about 1600 ng-hr/mL, at least about 1700 ng-hr/mL, at least about1800 ng-hr/mL, at least about 1900 ng-hr/mL, at least about 2000ng-hr/mL, at least about 2250 ng-hr/mL, or at least about 2500 ng-hr/mL.In particular embodiments, the AUC determination is obtained from atime-concentration pharmacokinetic profile obtained from the bloodsamples of animals or human volunteers following dosing.

Particular embodiments herein provide pharmaceutical formulations (e.g.,immediate release oral formulations and/or formulations that release theAPI substantially in the stomach) comprising a cytidine analog (e.g.,5-azacytidine or another cytidine analog provided herein) that achieve aparticular maximum plasma concentration (“Cmax”) in the subject to whichthe formulation is orally administered. Particular embodiments provideoral formulations that achieve a Cmax of the cytidine analog of at leastabout 25 ng/mL, at least about 50 ng/mL, at least about 75 ng/mL, atleast about 100 ng/mL, at least about 150 ng/mL, at least about 200ng/mL, at least about 250 ng/mL, at least about 300 ng/mL, at leastabout 350 ng/mL, at least about 400 ng/mL, at least about 450 ng/mL, atleast about 500 ng/mL, at least about 550 ng/mL, at least about 600ng/mL, at least about 650 ng/mL, at least about 700 ng/mL, at leastabout 750 ng/mL, at least about 800 ng/mL, at least about 850 ng/mL, atleast about 900 ng/mL, at least about 950 ng/mL, at least about 1000ng/mL, at least about 1100 ng/mL, at least about 1200 ng/mL, at leastabout 1300 ng/mL, at least about 1400 ng/mL, at least about 1500 ng/mL,at least about 1600 ng/mL, at least about 1700 ng/mL, at least about1800 ng/mL, at least about 1900 ng/mL, at least about 2000 ng/mL, atleast about 2250 ng/mL, or at least about 2500 ng/mL.

Particular embodiments herein provide pharmaceutical formulations (e.g.,immediate release oral formulations and/or formulations that release theAPI substantially in the stomach) comprising a cytidine analog (e.g.,5-azacytidine or another cytidine analog provided herein) that achieve aparticular time to maximum plasma concentration (“Tmax”) in the subjectto which the formulation is orally administered. Particular embodimentsprovide oral formulations that achieve a Tmax of the cytidine analog ofless than about 10 min., less than about 15 min., less than about 20min., less than about 25 min., less than about 30 min., less than about35 min., less than about 40 min., less than about 45 min., less thanabout 50 min., less than about 55 min., less than about 60 min., lessthan about 65 min., less than about 70 min., less than about 75 min.,less than about 80 min., less than about 85 min., less than about 90min., less than about 95 min., less than about 100 min., less than about105 min., less than about 110 min., less than about 115 min., less thanabout 120 min., less than about 130 min., less than about 140 min., lessthan about 150 min., less than about 160 min., less than about 170 min.,less than about 180 min., less than about 190 min., less than about 200min., less than about 210 min., less than about 220 min., less thanabout 230 min., or less than about 240 min. In particular embodiments,the Tmax value is measured from the time at which the formulation isorally administered.

Particular embodiments herein provide oral dosage forms comprising acytidine analog, wherein the oral dosage forms have an enteric coating.Particular embodiments provide a permeable or partly permeable (e.g.,“leaky”) enteric coating with pores. In particular embodiments, thepermeable or partly permeable enteric-coated tablet releases the5-azacytidine in an immediate release manner substantially in thestomach.

3. Compositions of Certain Dosage Forms Provided Herein

Provided herein are dosage forms designed to maximize the absorptionand/or efficacious delivery of certain cytidine analogs, e.g.,5-azacytidine or other cytidine analogs provided herein, upon oraladministration, e.g., for release substantially in the stomach.Accordingly, certain embodiments herein provide a solid oral dosage formof a cytidine analog, such as, for example, 5-azacytidine or anothercytidine analog provided herein, using pharmaceutical excipientsdesigned for immediate release of the API upon oral administration,e.g., substantially in the stomach. Particular immediate releaseformulations comprise a specific amount of a cytidine analog andoptionally one or more excipients. In certain embodiments, theformulation may be an immediate release tablet or an immediate releasecapsule (such as, e.g., an HPMC capsule).

Provided herein are methods of making the formulations provided hereincomprising a cytidine analog provided herein (e.g., immediate releaseoral formulations and/or formulations that release the API substantiallyin the stomach). In particular embodiments, the formulations providedherein may be prepared using conventional methods known to those skilledin the field of pharmaceutical formulation, as described, e.g., inpertinent textbooks. See, e.g., REMINGTON, THE SCIENCE AND PRACTICE OFPHARMACY, 20th Edition, Lippincott Williams & Wilkins, (2000); ANSEL etal., PHARMACEUTICAL DOSAGE FORMS AND DRUG DELIVERY SYSTEMS, 7th Edition,Lippincott Williams & Wilkins, (1999); GIBSON, PHARMACEUTICALPREFORMULATION AND FORMULATION, CRC Press (2001).

In particular embodiments, formulations provided herein (e.g., immediaterelease oral formulations, formulations that release the APIsubstantially in the stomach, or rapidly disintegrating formulationsthat dissolve substantially in the mouth) comprise a cytidine analog,such as, for example, 5-azacytidine or another cytidine analog providedherein, in a specific amount. In particular embodiments, the specificamount of the cytidine analog in the formulation is, e.g., about 10 mg,about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg,about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about95 mg, about 100 mg, about 120 mg, about 140 mg, about 150 mg, about 160mg, about 180 mg, about 200 mg, about 220 mg, least about 240 mg, about250 mg, about 260 mg, about 280 mg, about 300 mg, about 320 mg, about340 mg, about 350 mg, about 360 mg, about 380 mg, about 400 mg, about420 mg, about 440 mg, about 450 mg, about 460 mg, about 480 mg, about500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about1000 mg, about 1100 mg, about 1200 mg, about 1300 mg, about 1400 mg,about 1500 mg, about 1600 mg, about 1700 mg, about 1800 mg, about 1900mg, about 2000 mg, about 2100 mg, about 2200 mg, about 2300 mg, about2400 mg, about 2500 mg, about 3000 mg, about 4000 mg, or about 5000 mg.In particular embodiments, the specific amount of the cytidine analog inthe formulation is, e.g., at least about 10 mg, at least about 20 mg, atleast about 40 mg, at least about 60 mg, at least about 80 mg, at leastabout 100 mg, at least about 120 mg, at least about 140 mg, at leastabout 160 mg, at least about 180 mg, at least about 200 mg, at leastabout 220 mg, at least about 240 mg, at least about 250 mg, at leastabout 260 mg, at least about 280 mg, at least about 300 mg, at leastabout 320 mg, at least about 340 mg, at least about 350 mg, at leastabout 360 mg, at least about 380 mg, at least about 400 mg, at leastabout 420 mg, at least about 440 mg, at least about 450 mg, at leastabout 460 mg, at least about 480 mg, at least about 500 mg, at leastabout 550 mg, at least about 600 mg, at least about 650 mg, at leastabout 700 mg, at least about 750 mg, at least about 800 mg, at leastabout 900 mg, at least about 1000 mg, at least about 1100 mg, at leastabout 1200 mg, at least about 1300 mg, at least about 1400 mg, at leastabout 1500 mg, at least about 1600 mg, at least about 1700 mg, at leastabout 1800 mg, at least about 1900 mg, at least about 2000 mg, at leastabout 2100 mg, at least about 2200 mg, at least about 2300 mg, at leastabout 2400 mg, at least about 2500 mg, at least about 3000 mg, at leastabout 4000 mg, or at least about 5000 mg.

In certain embodiments, the formulation is a tablet, wherein the tabletis manufactured using standard, art-recognized tablet processingprocedures and equipment. In certain embodiments, the method for formingthe tablets is direct compression of a powdered, crystalline and/orgranular composition comprising the cytidine analog, alone or incombination with one or more excipients, such as, for example, carriers,additives, polymers, or the like. In certain embodiments, as analternative to direct compression, the tablets may be prepared using wetgranulation or dry granulation processes. In certain embodiments, thetablets are molded rather than compressed, starting with a moist orotherwise tractable material. In certain embodiments, compression andgranulation techniques are used.

In certain embodiments, the formulation is a capsule, wherein thecapsules may be manufactured using standard, art-recognized capsuleprocessing procedures and equipments. In certain embodiments, softgelatin capsules may be prepared in which the capsules contain a mixtureof the cytidine analog and vegetable oil or non-aqueous, water misciblematerials such as, for example, polyethylene glycol and the like. Incertain embodiments, hard gelatin capsules may be prepared containinggranules of the cytidine analog in combination with a solid pulverulentcarrier, such as, for example, lactose, saccharose, sorbitol, mannitol,potato starch, corn starch, amylopectin, cellulose derivatives, orgelatin. In certain embodiments, a hard gelatin capsule shell may beprepared from a capsule composition comprising gelatin and a smallamount of plasticizer such as glycerol. In certain embodiments, as analternative to gelatin, the capsule shell may be made of a carbohydratematerial. In certain embodiments, the capsule composition mayadditionally include polymers, colorings, flavorings and opacifiers asrequired. In certain embodiments, the capsule comprises HPMC.

In certain embodiments, the formulation of the cytidine analog, such as,for example, 5-azacytidine or another cytidine analog provided herein,is prepared using aqueous solvents without causing significanthydrolytic degradation of the cytidine analog. In particularembodiments, the formulation of the cytidine analog, such as, forexample, 5-azacytidine or another cytidine analog provided herein, is atablet which contains a coating applied to the drug core using aqueoussolvents without causing significant hydrolytic degradation of thecytidine analog in the formulation. In certain embodiments, water isemployed as the solvent for coating the drug core. In certainembodiments, the oral dosage form of the cytidine analog is a tabletcontaining a film coat applied to the drug core using aqueous solvents.In particular embodiments, water is employed as the solvent forfilm-coating. In particular embodiments, the tablet containing thecytidine analog is film-coated using aqueous solvents without effectingdegradation of the pharmaceutical composition. In particularembodiments, water is used as the film coating solvent without effectingdegradation of the pharmaceutical composition. In particularembodiments, an oral dosage form comprising 5-azacytidine and an aqueousfilm coating effects immediate drug release upon oral delivery. Inparticular embodiments, the oral dosage form comprising 5-azacytidineand an aqueous film coating effects controlled drug release to the uppergastrointestinal tract, e.g., the stomach, upon oral administration. Inparticular embodiments, a tablet with an aqueous-based film coatingcomprises 5-azacytidine as the API.

In certain embodiments, provided herein is a controlled releasepharmaceutical formulation for oral administration of a cytidine analogthat releases the cytidine analog substantially in the stomach,comprising: a) a specific amount of a cytidine analog; b) a drug releasecontrolling component for controlling the release of the cytidine analogsubstantially in the upper gastrointestinal tract, e.g., the stomach;and c) optionally one or more excipients. In certain embodiments, theoral dosage form comprising the cytidine analog is prepared as acontrolled release tablet or capsule which includes a drug corecomprising the pharmaceutical composition and optional excipients.Optionally, a “seal coat” or “shell” is applied. In certain embodiments,a formulation provided herein comprising a cytidine analog providedherein is a controlled release tablet or capsule, which comprises atherapeutically effective amount of the cytidine analog, a drug releasecontrolling component that controls the release of the cytidine analogsubstantially in the stomach upon oral administration, and optionally,one or more excipients.

Particular embodiments provide a drug release controlling component thatis a polymer matrix, which swells upon exposure to gastric fluid toeffect the gastric retention of the formulation and the sustainedrelease of the cytidine analog from the polymer matrix substantially inthe stomach. In certain embodiments, such formulations may be preparedby incorporating the cytidine analog into a suitable polymeric matrixduring formulation. Examples of such formulations are known in the art.See, e.g., Shell et al., U.S. Patent Publication No. 2002/0051820(application Ser. No. 09/990,061); Shell et al., U.S. Patent PublicationNo. 2003/0039688 (application Ser. No. 10/045,823); Gusler et al., U.S.Patent Publication No. 2003/0104053 (application Ser. No. 10/029,134),each of which is incorporated herein by reference in its entirety.

In certain embodiments, the drug release controlling component maycomprise a shell surrounding the drug-containing core, wherein the shellreleases the cytidine analog from the core by, e.g., permittingdiffusion of the cytidine analog from the core and promoting gastricretention of the formulation by swelling upon exposure to gastric fluidsto a size that is retained in the stomach. In certain embodiments, suchformulations may be prepared by first compressing a mixture of thecytidine analog and one or more excipients to form a drug core, andcompressing another powdered mixture over the drug core to form theshell, or enclosing the drug core with a capsule shell made of suitablematerials. Examples of such formulations are known in the art. See,e.g., Berner et al., U.S. Patent Publication No. 2003/0104062application Ser. No. 10/213,823), incorporated herein by reference inits entirety.

Certain embodiments herein provide oral dosage forms comprising acytidine analog, wherein the dosage form contains pores in theconventional enteric coating. In particular embodiments, the oral dosageform of the cytidine analog is a tablet that contains a permeable orpartly permeable (e.g., “leaky”) enteric coating with pores. Inparticular embodiments, the permeable or partly permeable enteric-coatedtablet controls the release of the cytidine analog from the tabletprimarily to the upper gastrointestinal tract, e.g., the stomach. Inparticular embodiments, the permeable or partly permeable enteric-coatedtablet comprises 5-azacytidine. In particular embodiments, the remainderof the cytidine analog is subsequently released beyond the stomach(e.g., in the intestine).

In certain embodiments, the pharmaceutical formulation provided hereinis a compressed tablet comprising a cytidine analog. In addition to thecytidine analog, the tablet optionally comprises one or more excipients,including (a) diluents or fillers, which may add necessary bulk to aformulation to prepare tablets of the desired size; (b) binders oradhesives, which may promote adhesion of the particles of theformulation, enabling a granulation to be prepared and maintaining theintegrity of the final tablet; (c) disintegrants or disintegratingagents, which, after administration, may promote breakup of the tabletsto smaller particles for improved drug availability; (d) anti-adherents,glidants, lubricants or lubricating agents, which may enhance flow ofthe tableting material into the tablet dies, minimize wear of thepunches and dies, prevent the sticking of fill material to the punchesand dies, and produce tablets having a sheen; and (e) miscellaneousadjuncts such as colorants and flavorants. After compression, tabletsprovided herein may be coated with various materials as describedherein.

In certain embodiments, the pharmaceutical formulation provided hereinis a multiple compressed tablet of a cytidine analog. Multiplecompressed tablets are prepared by subjecting the fill material to morethan a single compression. The result may be a multiple-layered tabletor a tablet-within-a-tablet, the inner tablet being the core comprisinga cytidine analog and optionally one or more excipients, and the outerportion being the shell, wherein the shell comprises one or moreexcipients, and may or may not contain the cytidine analog. Layeredtablets may be prepared by the initial compaction of a portion of fillmaterial in a die followed by additional fill material and compressionto form two- or three-layered tablets, depending upon the number ofseparate fills. Each layer may contain a different therapeutic agent,separate from one another for reasons of chemical or physicalincompatibility, or the same therapeutic agent for staged drug release,or simply for the unique appearance of the multiple-layered tablet. Eachportion of fill may be colored differently to prepare a distinctivelooking tablet. In the preparation of tablets having a compressed tabletas the inner core, special machines may be used to place the preformedtablet precisely within the die for the subsequent compression ofsurrounding fill material.

In certain embodiments, the compressed tablet of a cytidine analog maybe coated with a colored or an uncolored sugar layer. The coating may bewater-soluble and quickly dissolved after oral ingestion. The sugarcoating may serve the purpose of protecting the enclosed drug from theenvironment and providing a barrier to an objectionable taste or smell.The sugar coating may also enhance the appearance of the compressedtablet and permit the imprinting of identifying manufacturer'sinformation. In certain embodiments, sugar-coated tablets may be 50%larger and heavier than the original uncoated tablets. The sugar-coatingof tablets may be divided into the following optional steps: (1)waterproofing and sealing (if needed); (2) sub-coating; (3) smoothingand final rounding; (4) finishing and coloring (if desired); (5)imprinting (if needed); and (6) polishing.

In certain embodiments, the compressed tablet of a cytidine analog maybe film-coated. Film-coated tablets may be compressed tablets coatedwith a thin layer of a polymer capable of forming a skin-like film overthe tablet. The film is usually colored and has the advantage to be moredurable, less bulky, and less time-consuming to apply. By itscomposition, the coating may be designed to rupture and expose the coretablet at the desired location within the gastrointestinal tract. Thefilm-coating process, which places a thin skin-tight coating of aplastic-like material over the compressed tablet, may produce coatedtablets having essentially the same weight, shape, and size as theoriginally compressed tablet. The film-coating may be colored to makethe tablets attractive and distinctive. Film-coating solutions may benon-aqueous or aqueous. In particular embodiments, the non-aqueoussolutions may optionally contain one or more of the following types ofmaterials to provide the desired coating to the tablets: (1) a filmformer capable of producing smooth, thin films reproducible underconventional coating conditions and applicable to a variety of tabletshapes, such as, for example, cellulose acetate phthalate; (2) analloying substance providing water solubility or permeability to thefilm to ensure penetration by body fluids and therapeutic availabilityof the drug, such as, for example, polyethylene glycol; (3) aplasticizer to produce flexibility and elasticity of the coating andthus provide durability, such as, for example, castor oil; (4) asurfactant to enhance spreadability of the film during application, suchas, for example, polyoxyethylene sorbitan derivatives; (5) opaquants andcolorants to make the appearance of the coated tablets attractive anddistinctive, such as, for example, titanium dioxide as an opaquant, andFD&C or D&C dyes as a colorant; (6) sweeteners, flavors, or aromas toenhance the acceptability of the tablet to the subject, such as, forexample, saccharin as sweeteners, and vanillin as flavors and aromas;(7) a glossant to provide a luster to the tablets without a separatepolishing operation, such as, for example, beeswax; and (8) a volatilesolvent to allow the spread of the other components over the tabletswhile allowing rapid evaporation to permit an effective yet speedyoperation, such as, for example, alcohol-acetone mixture. In certainembodiments, an aqueous film-coating formulation may contain one or moreof the following: (1) film-forming polymer, such as, for example,cellulose ether polymers as hydroxypropyl methyl-cellulose,hydroxypropyl cellulose, and methyl-cellulose; (2) plasticizer, such as,for example, glycerin, propylene glycol, polyethylene glycol, diethylphthalate, and dibutyl subacetate; (3) colorant and opacifier, such as,for example, FD&C or D&C lakes and iron oxide pigments; or (4) vehicle,such as, for example, water.

In certain embodiments, the compressed tablet of a cytidine analog maybe compression-coated. The coating material, in the form of agranulation or powder, may be compressed onto a tablet core of drug witha special tablet press.

In certain embodiments, the pharmaceutical formulation is agelatin-coated tablet comprising a cytidine analog. A gelatin-coatedtablet is a capsule-shaped compressed tablet that allows the coatedproduct to be smaller than a capsule filled with an equivalent amount ofpowder. The gelatin coating facilitates swallowing and compared tounsealed capsules, gelatin-coated tablets may be more tamper-evident.

In certain embodiments, the pharmaceutical formulation may be asublingual tablet of a cytidine analog. The sublingual tablet isintended to be dissolved beneath the tongue for absorption through theoral mucosa. The sublingual tablet may dissolve promptly and providerapid release of the drug.

In certain embodiments, the pharmaceutical formulation is an immediaterelease tablet of a cytidine analog. In certain embodiments, theimmediate release tablet is designed, e.g., to disintegrate and releasethe API absent of any special rate-controlling features, such as specialcoatings and other techniques. In certain embodiments, the formulationis a rapidly disintegrating tablet that, e.g., dissolves substantiallyin the mouth following administration. In certain embodiments, thepharmaceutical formulation is an extended release tablet of a cytidineanalog. In certain embodiments, the extended release tablet is designed,e.g., to release the API over an extended period of time andsubstantially in the stomach.

In certain embodiments, compressed tablets may be prepared by wetgranulation. Wet granulation is a widely employed method for theproduction of compressed tablets, and, in particular embodiments,requires one or more the following steps: (1) weighing and blending theingredients; (2) preparing a damp mass; (3) screening the damp mass intopellets or granules; (4) drying the granulation; (5) sizing thegranulation by dry screening; (6) adding lubricant and blending; and (7)tableting by compression.

In certain embodiments, compressed tablets may be prepared by drygranulation. By the dry granulation method, the powder mixture iscompacted in large pieces and subsequently broken down or sized intogranules. But this method, either the active ingredient or the diluenthas cohesive property. After weighing and mixing the ingredients, thepowder mixture may be slugged or compressed into large flat tablets orpellets. The slugs then are broken up by hand or by a mill and passedthrough a screen of desired mesh for sizing. Lubricant is added in theusual manner, and tablets are prepared by compression. Alternatively,instead of slugging, powder compactors may be used to increase thedensity of a powder by pressing it between high-pressure rollers. Thecompressed material then is broken up, sized, and lubricated, andtablets are prepared by compression in the usual manner. The rollercompaction method is often preferred over slugging. Binding agents usedin roller compaction formulations include methylcellulose orhydroxyl-methylcellulose and can produce good tablet hardness andfriability.

In certain embodiments, compressed tablets may be prepared by directcompression. Some granular chemicals possess free flowing and cohesiveproperties that enable them to be compressed directly in a tabletmachine without the need of wet or dry granulation. For chemicals thatdo not possess this quality, special pharmaceutical excipients may beused which impart the necessary qualities for the production of tabletsby direct compression. Particular tableting excipients include, e.g.:fillers, such as spray-dried lactose, micro-crystals ofalpha-monohydrate lactose, sucrose-invert sugar-corn starch mixtures,micro-crystalline cellulose, crystalline maltose, and di-calciumphosphate; disintegrating agents, such as direct-compression starch,sodium carboxymethyl starch, cross-linked carboxymethylcellulose fibers,and cross-linked polyvinylpyrrolidone; lubricants, such as magnesiumsearate and talc; and glidants, such as fumed silicon dioxide.

In certain embodiments, tablets provided herein may be prepared bymolding. The base for molded tablets is generally a mixture of finelypowdered lactose with or without a portion of powdered sucrose. Inpreparing the fill, the drug is mixed uniformly with the base bygeometric dilution. The powder mixture may be wetted with a mixture ofwater and alcohol sufficient only to dampen the powder so that it may becompacted. The solvent action of the water on a portion of thelactose/sucrose base effects the biding of the powder mixture upondrying. The alcohol portion hastens the drying process.

In certain embodiments, the pharmaceutical formulations provided hereincontain a cytidine analog and, optionally, one or more excipients toform a “drug core.” Optional excipients include, e.g., diluents (bulkingagents), lubricants, disintegrants, fillers, stabilizers, surfactants,preservatives, coloring agents, flavoring agents, binding agents,excipient supports, glidants, permeation enhancement excipients,plasticizers and the like, e.g., as known in the art. It will beunderstood by those in the art that some substances serve more than onepurpose in a pharmaceutical composition. For instance, some substancesare binders that help hold a tablet together after compression, yet arealso disintegrants that help break the tablet apart once it reaches thetarget delivery site. Selection of excipients and amounts to use may bereadily determined by the formulation scientist based upon experienceand consideration of standard procedures and reference works availablein the art.

In certain embodiments, formulations provided herein comprise one ormore binders. Binders may be used, e.g., to impart cohesive qualities toa tablet, and thus ensure that the tablet remains intact aftercompression. Suitable binders include, but are not limited to, starch(including corn starch and pregelatinized starch), gelatin, sugars(including sucrose, glucose, dextrose and lactose), polyethylene glycol,propylene glycol, waxes, and natural and synthetic gums, e.g., acaciasodium alginate, polyvinylpyrrolidone, cellulosic polymers (includinghydroxypropyl cellulose, hydroxypropylmethylcellulose, methyl cellulose,ethyl cellulose, hydroxyethyl cellulose, carboxymethyl cellulose and thelike), veegum, carbomer (e.g., carbopol), sodium, dextrin, guar gum,hydrogenated vegetable oil, magnesium aluminum silicate, maltodextrin,polymethacrylates, povidone (e.g., KOLLIDON, PLASDONE), microcrystallinecellulose, among others. Binding agents also include, e.g., acacia,agar, alginic acid, cabomers, carrageenan, cellulose acetate phthalate,ceratonia, chitosan, confectioner's sugar, copovidone, dextrates,dextrin, dextrose, ethylcellulose, gelatin, glyceryl behenate, guar gum,hydroxyethyl cellulose, hydroxyethylmethyl cellulose, hydroxypropylcellulose, hydroxypropyl starch, hypromellose, inulin, lactose,magnesium aluminum silicate, maltodextrin, maltose, methylcellulose,poloxamer, polycarbophil, polydextrose, polyethylene oxide,polymethylacrylates, povidone, sodium alginate, sodiumcarboxymethylcellulose, starch, pregelatinized starch, stearic acid,sucrose, and zein. The binding agent can be, relative to the drug core,in the amount of about 2% w/w of the drug core; about 4% w/w of the drugcore, about 6% w/w of the drug core, about 8% w/w of the drug core,about 10% w/w of the drug core, about 12% w/w of the drug core, about14% w/w of the drug core, about 16% w/w of the drug core, about 18% w/wof the drug core, about 20% w/w of the drug core, about 22% w/w of thedrug core, about 24% w/w of the drug core, about 26% w/w of the drugcore, about 28% w/w of the drug core, about 30% w/w of the drug core,about 32% w/w of the drug core, about 34% w/w of the drug core, about36% w/w of the drug core, about 38% w/w of the drug core, about 40% w/wof the drug core, about 42% w/w of the drug core, about 44% w/w of thedrug core, about 46% w/w of the drug core, about 48% w/w of the drugcore, about 50% w/w of the drug core, about 52% w/w of the drug core,about 54% w/w of the drug core, about 56% w/w of the drug core, about58% w/w of the drug core, about 60% w/w of the drug core, about 62% w/wof the drug core, about 64% w/w of the drug core, about 66% w/w of thedrug core; about 68% w/w of the drug core, about 70% w/w of the drugcore, about 72% w/w of the drug core, about 74% w/w of the drug core,about 76% w/w of the drug core, about 78% w/w of the drug core, about80% w/w of the drug core, about 82% w/w of the drug core, about 84% w/wof the drug core, about 86% w/w of the drug core, about 88% w/w of thedrug core, about 90% w/w of the drug core, about 92% w/w of the drugcore, about 94% w/w of the drug core, about 96% w/w of the drug core,about 98% w/w of the drug core, or more, if determined to beappropriate. In certain embodiments, a suitable amount of a particularbinder is determined by one of ordinary skill in the art.

In certain embodiments, formulations provided herein comprise one ormore diluents. Diluents may be used, e.g., to increase bulk so that apractical size tablet is ultimately provided. Suitable diluents includedicalcium phosphate, calcium sulfate, lactose, cellulose, kaolin,mannitol, sodium chloride, dry starch, microcrystalline cellulose (e.g.,AVICEL), microfine cellulose, pregelitinized starch, calcium carbonate,calcium sulfate, sugar, dextrates, dextrin, dextrose, dibasic calciumphosphate dihydrate, tribasic calcium phosphate, kaolin, magnesiumcarbonate, magnesium oxide, maltodextrin, mannitol, polymethacrylates(e.g., EUDRAGIT), potassium chloride, sodium chloride, sorbitol andtalc, among others. Diluents also include, e.g., ammonium alginate,calcium carbonate, calcium phosphate, calcium sulfate, celluloseacetate, compressible sugar, confectioner's sugar, dextrates, dextrin,dextrose, erythritol, ethylcellulose, fructose, fumaric acid, glycerylpalmitostearate, isomalt, kaolin, lacitol, lactose, mannitol, magnesiumcarbonate, magnesium oxide, maltodextrin, maltose, medium-chaintriglycerides, microcrystalline cellulose, microcrystalline silicifiedcellulose, powered cellulose, polydextrose, polymethylacrylates,simethicone, sodium alginate, sodium chloride, sorbitol, starch,pregelatinized starch, sucrose, sulfobutylether-β-cyclodextrin, talc,tragacanth, trehalose, and xylitol. Diluents may be used in amountscalculated to obtain a desired volume for a tablet or capsule; incertain embodiments, a diluent is used in an amount of about 5% or more,about 10% or more, about 15% or more, about 20% or more, about 22% ormore, about 24% or more, about 26% or more, about 28% or more, about 30%or more, about 32% or more, about 34% or more, about 36% or more, about38% or more, about 40% or more, about 42% or more, about 44% or more,about 46% or more, about 48% or more, about 50% or more, about 52% ormore, about 54% or more, about 56% or more, about 58% or more, about 60%or more, about 62% or more, about 64% or more, about 68% or more, about70% or more, about 72% or more, about 74% or more, about 76% or more,about 78% or more, about 80% or more, about 85% or more, about 90% ormore, or about 95% or more, weight/weight, of a drug core; between about10% and about 90% w/w of the drug core; between about 20% and about 80%w/w of the drug core; between about 30% and about 70% w/w of the drugcore; between about 40% and about 60% w/w of the drug core. In certainembodiments, a suitable amount of a particular diluent is determined byone of ordinary skill in the art.

In certain embodiments, formulations provided herein comprise one ormore lubricants. Lubricants may be used, e.g., to facilitate tabletmanufacture; examples of suitable lubricants include, for example,vegetable oils such as peanut oil, cottonseed oil, sesame oil, oliveoil, corn oil, and oil of theobroma, glycerin, magnesium stearate,calcium stearate, and stearic acid. In certain embodiments, stearates,if present, represent no more than approximately 2 weight % of thedrug-containing core. Further examples of lubricants include, e.g.,calcium stearate, glycerin monostearate, glyceryl behenate, glycerylpalmitostearate, magnesium lauryl sulfate, magnesium stearate, myristicacid, palmitic acid, poloxamer, polyethylene glycol, potassium benzoate,sodium benzoate, sodium chloride, sodium lauryl sulfate, sodium stearylfumarate, stearic acid, talc, and zinc stearate. In particularembodiments, the lubricant is magnesium stearate. In certainembodiments, the lubricant is present, relative to the drug core, in anamount of about 0.2% w/w of the drug core, about 0.4% w/w of the drugcore, about 0.6% w/w of the drug core, about 0.8% w/w of the drug core,about 1.0% w/w of the drug core, about 1.2% w/w of the drug core, about1.4% w/w of the drug core, about 1.6% w/w of the drug core, about 1.8%w/w of the drug core, about 2.0% w/w of the drug core, about 2.2% w/w ofthe drug core, about 2.4% w/w of the drug core, about 2.6% w/w of thedrug core, about 2.8% w/w of the drug core, about 3.0% w/w of the drugcore, about 3.5% w/w of the drug core, about 4% w/w of the drug core,about 4.5% w/w of the drug core, about 5% w/w of the drug core, about 6%w/w of the drug core, about 7% w/w of the drug core, about 8% w/w of thedrug core, about 10% w/w of the drug core, about 12% w/w of the drugcore, about 14% w/w of the drug core, about 16% w/w of the drug core,about 18% w/w of the drug core, about 20% w/w of the drug core, about25% w/w of the drug core, about 30% w/w of the drug core, about 35% w/wof the drug core, about 40% w/w of the drug core, between about 0.2% andabout 10% w/w of the drug core, between about 0.5% and about 5% w/w ofthe drug core, or between about 1% and about 3% w/w of the drug core. Incertain embodiments, a suitable amount of a particular lubricant isdetermined by one of ordinary skill in the art.

In certain embodiments, formulations provided herein comprise one ormore disintegrants. Disintegrants may be used, e.g., to facilitatedisintegration of the tablet, and may be, e.g., starches, clays,celluloses, algins, gums or crosslinked polymers. Disintegrants alsoinclude, e.g., alginic acid, carboxymethylcellulose calcium,carboxymethylcellulose sodium (e.g., AC-DI-SOL, PRIMELLOSE), colloidalsilicon dioxide, croscarmellose sodium, crospovidone (e.g., KOLLIDON,POLYPLASDONE), guar gum, magnesium aluminum silicate, methyl cellulose,microcrystalline cellulose, polacrilin potassium, powdered cellulose,pregelatinized starch, sodium alginate, sodium starch glycolate (e.g.,EXPLOTAB) and starch. Additional disintegrants include, e.g., calciumalginate, chitosan, sodium docusate, hydroxypropyl cellulose, andpovidone. In certain embodiments, the disintegrant is, relative to thedrug core, present in the amount of about 1% w/w of the drug core, about2% w/w of the drug core, about 3% w/w of the drug core, about 4% w/w ofthe drug core, about 5% w/w of the drug core, about 6% w/w of the drugcore, about 7% w/w of the drug core, about 8% w/w of the drug core,about 9% w/w of the drug core, about 10% w/w of the drug core, about 12%w/w of the drug core, about 14% w/w of the drug core, about 16% w/w ofthe drug core, about 18% w/w of the drug core, about 20% w/w of the drugcore, about 22% w/w of the drug core, about 24% w/w of the drug core,about 26% w/w of the drug core, about 28% w/w of the drug core, about30% w/w of the drug core, about 32% w/w of the drug core, greater thanabout 32% w/w of the drug core, between about 1% and about 10% w/w ofthe drug core, between about 2% and about 8% w/w of the drug core,between about 3% and about 7% w/w of the drug core, or between about 4%and about 6% w/w of the drug core. In certain embodiments, a suitableamount of a particular disintegrant is determined by one of ordinaryskill in the art.

In certain embodiments, formulations provided herein comprise one ormore stabilizers. Stabilizers (also called absorption enhancers) may beused, e.g., to inhibit or retard drug decomposition reactions thatinclude, by way of example, oxidative reactions. Stabilizing agentsinclude, e.g., d-Alpha-tocopheryl polyethylene glycol 1000 succinate(Vitamin E TPGS), acacia, albumin, alginic acid, aluminum stearate,ammonium alginate, ascorbic acid, ascorbyl palmitate, bentonite,butylated hydroxytoluene, calcium alginate, calcium stearate, calciumcarboxymethylcellulose, carrageenan, ceratonia, colloidal silicondioxide, cyclodextrins, diethanolamine, edetates, ethylcellulose,ethyleneglycol palmitostearate, glycerin monostearate, guar gum,hydroxypropyl cellulose, hypromellose, invert sugar, lecithin, magnesiumaluminum silicate, monoethanolamine, pectin, poloxamer, polyvinylalcohol, potassium alginate, potassium polacrilin, povidone, propylgallate, propylene glycol, propylene glycol alginate, raffinose, sodiumacetate, sodium alginate, sodium borate, sodium carboxymethyl cellulose,sodium stearyl fumarate, sorbitol, stearyl alcohol,sufobutyl-b-cyclodextrin, trehalose, white wax, xanthan gum, xylitol,yellow wax, and zinc acetate. In certain embodiments, the stabilizer is,relative to the drug core, present in the amount of about 1% w/w of thedrug core, about 2% w/w of the drug core, about 3% w/w of the drug core,about 4% w/w of the drug core, about 5% w/w of the drug core, about 6%w/w of the drug core, about 7% w/w of the drug core, about 8% w/w of thedrug core, about 9% w/w of the drug core, about 10% w/w of the drugcore, about 12% w/w of the drug core, about 14% w/w of the drug core,about 16% w/w of the drug core, about 18% w/w of the drug core, about20% w/w of the drug core, about 22% w/w of the drug core, about 24% w/wof the drug core, about 26% w/w of the drug core, about 28% w/w of thedrug core, about 30% w/w of the drug core, about 32% w/w of the drugcore, between about 1% and about 10% w/w of the drug core, between about2% and about 8% w/w of the drug core, between about 3% and about 7% w/wof the drug core, or between about 4% and about 6% w/w of the drug core.In certain embodiments, a suitable amount of a particular stabilizer isdetermined by one of ordinary skill in the art.

In certain embodiments, formulations provided herein comprise one ormore glidants. Glidants may be used, e.g., to improve the flowproperties of a powder composition or granulate or to improve theaccuracy of dosing. Excipients that may function as glidants include,e.g., colloidal silicon dioxide, magnesium trisilicate, powderedcellulose, starch, tribasic calcium phosphate, calcium silicate,powdered cellulose, colloidal silicon dioxide, magnesium silicate,magnesium trisilicate, silicon dioxide, starch, tribasic calciumphosphate, and talc. In certain embodiments, the glidant is, relative tothe drug core, present in the amount of less than about 1% w/w of thedrug core, about 1% w/w of the drug core, about 2% w/w of the drug core,about 3% w/w of the drug core, about 4% w/w of the drug core, about 5%w/w of the drug core, about 6% w/w of the drug core, about 7% w/w of thedrug core, about 8% w/w of the drug core, about 9% w/w of the drug core,about 10% w/w of the drug core, about 12% w/w of the drug core, about14% w/w of the drug core, about 16% w/w of the drug core, about 18% w/wof the drug core, about 20% w/w of the drug core, about 22% w/w of thedrug core, about 24% w/w of the drug core, about 26% w/w of the drugcore, about 28% w/w of the drug core, about 30% w/w of the drug core,about 32% w/w of the drug core, between about 1% and about 10% w/w ofthe drug core, between about 2% and about 8% w/w of the drug core,between about 3% and about 7% w/w of the drug core, or between about 4%and about 6% w/w of the drug core. In certain embodiments, a suitableamount of a particular glidant is determined by one of ordinary skill inthe art.

In one embodiment, formulations provided herein comprise one or morecomplexing agents. In certain embodiments, the complexing agentsinclude, but are not limited to, cyclodextrins, includingα-cyclodextrin, β-cyclodextrin, hydroxypropyl-β-cyclodextrin,sulfobutylether-β-cyclodextrin, and sulfobutylether 7-β-cyclodextrin(CAPTISOL®, CyDex, Lenexa, Kans.).

In certain embodiments, formulations provided herein comprise one ormore permeation enhancers (also called, e.g., permeability enhancers).In certain embodiments, the permeation enhancer enhances the uptake of acytidine analog through the gastrointestinal wall (e.g., the stomach).In certain embodiments, the permeation enhancer alters the rate and/oramount of the cytidine analog that enters the bloodstream. In particularembodiments, d-alpha-tocopheryl polyethylene glycol-1000 succinate(Vitamin E TPGS) is used as a permeation enhancer. In particularembodiments, one or more other suitable permeation enhancers are used,including, e.g., any permeation enhancer known in the art. Specificexamples of suitable permeation enhancers include, e.g., those listedbelow:

Example of Product name Chemical Name Supplier Pluronic F 127 PoloxamerF 127 Sigma Lutrol F 68 Poloxamer 188 BASF Carbopol 934-P Carbomer 934-PSpectrum Chemical Tween 80 Polysorbate 80 Sigma Chitosan Chitosan LowMol Wt Aldrich Capric acid/Na cap Sodium Decanoate Sigma Lauric acid/Nalaur Sodium Dodecanoate Sigma Disodium EDTA Ethylenediamine tetraaceticSigma acid disodium dehydrate Propylene glycol 1,2 Propanediol Sigma CMCellulose Carboxymethyl Cellulose Sigma Labrasol Caprylocaproylmacrogol-8 Gattefosse glycerides N,N-Dimethylacetamide (minimum 99%)Sigma Vitamin E TPGS d-Alpha-Tocopheryl Eastman Polyethylene Glycol-1000Succinate Solutol HS 15 Polyethylene glycol 660 BASF 12-hydroxystearateLabrafil M 1944 CS (2) Oleyl Macrogolglyerides Gattefosse

Other potential permeation enhancers include, e.g., alcohols, dimethylsulfoxide, glyceryl monooleate, glycofurol, isopropyl myristate,isopropyl palmitate, lanolin, linoleic acid, myristic acid, oleic acid,oleyl alcohol, palmitic acid, polyoxyethylene alkyl ethers,2-pyrrolidone, sodium lauryl sulfate, and thymol.

In certain embodiments, the permeation enhancer is present in theformulation in an amount by weight, relative to the total weight of theformulation, of about 0.1%, about 0.2%, about 0.3%, about 0.4%, about0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about1.1%, about 1.2%, about 1.3%, about 1.4%, about 1.5%, about 1.6%, about1.7%, about 1.8%, about 1.9%, about 2%, about 2.1%, about 2.2%, about2.3%, about 2.4%, about 2.5%, about 2.6%, about 2.7%, about 2.8%, about2.9%, about 3%, about 3.1%, about 3.2%, about 3.3%, about 3.4%, about3.5%, about 3.6%, about 3.7%, about 3.8%, about 3.9%, about 4%, about4.1% about 4.2%, about 4.3%, about 4.4%, about 4.5%, about 4.6%, about4.7%, about 4.8%, about 4.9%, about 5%, about 5.1% about 5.2%, about5.3%, about 5.4%, about 5.5%, about 5.6%, about 5.7%, about 5.8%, about5.9%, about 6%, about 6.1% about 6.2%, about 6.3%, about 6.4%, about6.5%, about 6.6%, about 6.7%, about 6.8%, about 6.9%, about 7%, about7.1% about 7.2%, about 7.3%, about 7.4%, about 7.5%, about 7.6%, about7.7%, about 7.8%, about 7.9%, about 8%, about 8.1% about 8.2%, about8.3%, about 8.4%, about 8.5%, about 8.6%, about 8.7%, about 8.8%, about8.9%, about 9%, about 9.1% about 9.2%, about 9.3%, about 9.4%, about9.5%, about 9.6%, about 9.7%, about 9.8%, about 9.9%, about 10%, greaterthan about 10%, greater than about 12%, greater than about 14%, greaterthan about 16%, greater than about 18%, greater than about 20%, greaterthan about 25%, greater than about 30%, greater than about 35%, greaterthan about 40%, greater than about 45%, or greater than about 50%. Incertain embodiments, the appropriate amount of a suitable permeationenhancer provided herein is determined by one of skill in the art.

Without intending to be limited to any particular theory, the permeationenhancers provided herein may function by, inter alia, facilitating(e.g., increasing the rate or extent of) the transport of a cytidineanalog through the gastrointestinal wall. In general, movement throughthe gastrointestinal wall may occur by, e.g.: passive diffusion, such asthe movement of drug across a membrane in a manner driven solely by theconcentration gradient; carrier-mediated diffusion, such as the movementof drug across a cell membrane via a specialized transport systemembedded in the cell membrane; paracellular diffusion, such as themovement of a drug across a membrane by going between, rather thanthrough, two cells; and transcellular diffusion, such as the movement ofa drug across the cell. Additionally, there are numerous cellularproteins capable of preventing intracellular accumulation of drugs bypumping out drug that enters the cell. These are sometimes called effluxpumps. One such efflux pump is that involving p-glycoprotein, which ispresent in many different tissues in the body (e.g., intestine,placental membrane, blood-brain barrier). Permeation enhancers canfunction by, inter alia, facilitating any of the processes mentionedabove (such as by increasing fluidity of membranes, opening tightjunctions between cells, and/or inhibiting efflux, among others).

In certain embodiments, the compositions provided herein comprising acytidine analog, e.g., 5-azacytidine or another cytidine analog providedherein, are essentially free of a cytidine deaminase inhibitor (e.g., donot comprise a cytidine deaminase inhibitor). In certain embodiments,the compositions provided herein are essentially free of (e.g., do notcomprise) the cytidine deaminase inhibitor tetrahydrouridine (THU).Certain embodiments herein provide pharmaceutical compositionscomprising a therapeutically effective amount of a cytidine analog(e.g., 5-azacytidine or another cytidine analog provided herein),wherein the compositions release the cytidine analog substantially inthe stomach following oral administration to a subject, and wherein thecompositions are essentially free of (e.g., do not comprise) a cytidinedeaminase inhibitor (e.g., THU). Certain embodiments herein providepharmaceutical compositions comprising a therapeutically effectiveamount of a cytidine analog (e.g., 5-azacytidine or another cytidineanalog provided herein), wherein the compositions release the cytidineanalog substantially in the stomach following oral administration to asubject, wherein the compositions are essentially free of (e.g., do notcomprise) a cytidine deaminase inhibitor (e.g., THU), and wherein thecompositions achieve a particular biological parameter provided herein(e.g., a particular Cmax value, Tmax value, and/or AUC value providedherein). In particular embodiments, a composition provided herein thatis essentially free of a cytidine deaminase inhibitor (e.g., THU)comprises, e.g., less than 200 mg, less than 150 mg, less than 100 mg,less than 50 mg, less than 25 mg, less than 10 mg, less than 5 mg, lessthan 1 mg, or less than 0.1 mg of the cytidine deaminase inhibitor.

4. Other Embodiments of Oral Dosage Forms

In other embodiments, the pharmaceutical compositions provided hereinmay be provided as compressed tablets, tablet triturates, chewablelozenges, rapidly dissolving tablets, multiple compressed tablets, orenteric-coating tablets, sugar-coated, or film-coated tablets. In oneembodiment, enteric-coated tablets are compressed tablets coated withsubstances that resist the action of stomach acid but dissolve ordisintegrate in the intestine, thus protecting the active ingredientsfrom the acidic environment of the stomach. Enteric-coatings include,but are not limited to, fatty acids, fats, phenyl salicylate, waxes,shellac, ammoniated shellac, and cellulose acetate phthalates.Sugar-coated tablets are compressed tablets surrounded by a sugarcoating, which may be beneficial in covering up objectionable tastes orodors and in protecting the tablets from oxidation. Film-coated tabletsare compressed tablets that are covered with a thin layer or film of awater-soluble material. Film coatings include, but are not limited to,hydroxyethylcellulose, sodium carboxymethylcellulose, polyethyleneglycol 4000, and cellulose acetate phthalate. In one embodiment, filmcoating imparts the same general characteristics as sugar coating.Multiple compressed tablets are compressed tablets made by more than onecompression cycle, including layered tablets, and press-coated ordry-coated tablets.

In one embodiment, the tablet dosage forms may be prepared from theactive ingredient in powdered, crystalline, or granular forms, alone orin combination with one or more carriers or excipients described herein,including binders, disintegrants, controlled-release polymers,lubricants, diluents, and/or colorants. Flavoring and sweetening agentsare useful in the formation of chewable tablets and lozenges.

In one embodiment, the pharmaceutical compositions provided herein maybe provided as soft or hard capsules, which can be made from gelatin,methylcellulose, starch, or calcium alginate. The hard gelatin capsule,also known as the dry-filled capsule (DFC), consists of two sections,one slipping over the other, thus completely enclosing the activeingredient. The soft elastic capsule (SEC) is a soft, globular shell,such as a gelatin shell, which is plasticized by the addition ofglycerin, sorbitol, or a similar polyol. The soft gelatin shells maycontain a preservative to prevent the growth of microorganisms. Suitablepreservatives are those as described herein, including methyl- andpropyl-parabens, and sorbic acid. The liquid, semisolid, and soliddosage forms provided herein may be encapsulated in a capsule. Suitableliquid and semisolid dosage forms include solutions and suspensions inpropylene carbonate, vegetable oils, or triglycerides. Capsulescontaining such solutions can be prepared as described in U.S. Pat. Nos.4,328,245; 4,409,239; and 4,410,545. The capsules may also be coated asknown by those of skill in the art in order to modify or sustaindissolution of the active ingredient.

In one embodiment, the pharmaceutical compositions provided herein maybe provided in liquid and semisolid dosage forms, including emulsions,solutions, suspensions, elixirs, and syrups. An emulsion is a two-phasesystem, in which one liquid is dispersed in the form of small globulesthroughout another liquid, which can be oil-in-water or water-in-oil.Emulsions may include a pharmaceutically acceptable non-aqueous liquidor solvent, emulsifying agent, and preservative. Suspensions may includea pharmaceutically acceptable suspending agent and preservative. Aqueousalcoholic solutions may include a pharmaceutically acceptable acetal,such as a di(lower alkyl) acetal of a lower alkyl aldehyde, e.g.,acetaldehyde diethyl acetal; and a water-miscible solvent having one ormore hydroxyl groups, such as propylene glycol and ethanol. Elixirs areclear, sweetened, and hydroalcoholic solutions. Syrups are concentratedaqueous solutions of a sugar, for example, sucrose, and may also containa preservative. For a liquid dosage form, for example, a solution in apolyethylene glycol may be diluted with a sufficient quantity of apharmaceutically acceptable liquid carrier, e.g., water, to be measuredconveniently for administration.

In one embodiment, other useful liquid and semisolid dosage formsinclude, but are not limited to, those containing the activeingredient(s) provided herein, and a dialkylated mono- or poly-alkyleneglycol, including, 1,2-dimethoxymethane, diglyme, triglyme, tetraglyme,polyethylene glycol-350-dimethyl ether, polyethylene glycol-550-dimethylether, polyethylene glycol-750-dimethyl ether, wherein 350, 550, and 750refer to the approximate average molecular weight of the polyethyleneglycol. These formulations may further comprise one or moreantioxidants, such as butylated hydroxytoluene (BHT), butylatedhydroxyanisole (BHA), propyl gallate, vitamin E, hydroquinone,hydroxycoumarins, ethanolamine, lecithin, cephalin, ascorbic acid, malicacid, sorbitol, phosphoric acid, bisulfite, sodium metabisulfite,thiodipropionic acid and its esters, and dithiocarbamates.

In one embodiment, the pharmaceutical compositions provided herein maybe provided as non-effervescent or effervescent, granules and powders,to be reconstituted into a liquid dosage form. Pharmaceuticallyacceptable carriers and excipients used in the non-effervescent granulesor powders may include diluents, sweeteners, and wetting agents.Pharmaceutically acceptable carriers and excipients used in theeffervescent granules or powders may include organic acids and a sourceof carbon dioxide.

Coloring and flavoring agents can be used in the dosage forms providedherein.

In one embodiment, the pharmaceutical compositions provided herein maybe formulated as immediate or modified release dosage forms, includingdelayed-, sustained-, pulsed-, controlled-, targeted-, andprogrammed-release forms.

In one embodiment, the pharmaceutical compositions provided herein maybe co-formulated with other active ingredients which do not impair thedesired therapeutic action, or with substances that supplement thedesired action.

In one embodiment, active ingredients provided herein can beadministered by controlled release means or by delivery devices that arewell known to those of ordinary skill in the art. Examples include, butare not limited to, those described in U.S. Pat. Nos. 3,845,770;3,916,899; 3,536,809; 3,598,123; and 4,008,719, 5,674,533, 5,059,595,5,591,767, 5,120,548, 5,073,543, 5,639,476, 5,354,556, and 5,733,566,each of which is incorporated herein by reference. Such dosage forms canbe used to provide slow or controlled-release of one or more activeingredients using, for example, hydropropylmethyl cellulose, otherpolymer matrices, gels, permeable membranes, osmotic systems, multilayercoatings, microparticles, liposomes, microspheres, or a combinationthereof to provide the desired release profile in varying proportions.Suitable controlled-release formulations known to those of ordinaryskill in the art, including those described herein, can be readilyselected for use with the active agents provided herein. In oneembodiment, provided are single unit dosage forms suitable for oraladministration such as, but not limited to, tablets, capsules, gelcaps,and caplets that are adapted for controlled-release.

In one embodiment, controlled-release pharmaceutical products improvedrug therapy over that achieved by their non-controlled counterparts. Inanother embodiment, the use of a controlled-release preparation inmedical treatment is characterized by a minimum of drug substance beingemployed to cure or control the condition in a minimum amount of time.Advantages of controlled-release formulations include extended activityof the drug, reduced dosage frequency, and increased patient compliance.In addition, controlled-release formulations can be used to affect thetime of onset of action or other characteristics, such as blood levelsof the drug, and can thus affect the occurrence of side (e.g., adverse)effects.

In another embodiment, the controlled-release formulations are designedto initially release an amount of drug (active ingredient) that promptlyproduces the desired therapeutic or prophylactic effect, and graduallyand continually release of other amounts of drug to maintain this levelof therapeutic or prophylactic effect over an extended period of time.In one embodiment, in order to maintain a constant level of drug in thebody, the drug can be released from the dosage form at a rate that willreplace the amount of drug being metabolized and excreted from the body.Controlled-release of an active ingredient can be stimulated by variousconditions including, but not limited to, pH, temperature, enzymes,water, or other physiological conditions or compounds.

5. Parenteral Dosage Forms

In another embodiment, the pharmaceutical compositions provided hereinmay be administered parenterally by injection, infusion, orimplantation, for local or systemic administration. Parenteraladministration, as used herein, include intravenous, intraarterial,intraperitoneal, intrathecal, intraventricular, intraurethral,intrasternal, intracranial, intramuscular, intrasynovial, andsubcutaneous administration.

In one embodiment, the pharmaceutical compositions provided herein maybe formulated in dosage forms that are suitable for parenteraladministration, including solutions, suspensions, emulsions, micelles,liposomes, microspheres, nanosystems, and solid forms suitable forsolutions or suspensions in liquid prior to injection. Such dosage formscan be prepared according to conventional methods known to those skilledin the art of pharmaceutical science (see, e.g., Remington, The Scienceand Practice of Pharmacy, supra).

In one embodiment, the pharmaceutical compositions intended forparenteral administration may include one or more pharmaceuticallyacceptable carriers and excipients, including, but not limited to,aqueous vehicles, water-miscible vehicles, non-aqueous vehicles,antimicrobial agents or preservatives against the growth ofmicroorganisms, stabilizers, solubility enhancers, isotonic agents,buffering agents, antioxidants, local anesthetics, suspending anddispersing agents, wetting or emulsifying agents, complexing agents,sequestering or chelating agents, cryoprotectants, lyoprotectants,thickening agents, pH adjusting agents, and inert gases.

In one embodiment, suitable aqueous vehicles include, but are notlimited to, water, saline, physiological saline or phosphate bufferedsaline (PBS), sodium chloride injection, Ringers injection, isotonicdextrose injection, sterile water injection, dextrose and lactatedRingers injection. Non-aqueous vehicles include, but are not limited to,fixed oils of vegetable origin, castor oil, corn oil, cottonseed oil,olive oil, peanut oil, peppermint oil, safflower oil, sesame oil,soybean oil, hydrogenated vegetable oils, hydrogenated soybean oil, andmedium-chain triglycerides of coconut oil, and palm seed oil.Water-miscible vehicles include, but are not limited to, ethanol,1,3-butanediol, liquid polyethylene glycol (e.g., polyethylene glycol300 and polyethylene glycol 400), propylene glycol, glycerin,N-methyl-2-pyrrolidone, N,N-dimethylacetamide, and dimethyl sulfoxide.

In one embodiment, suitable antimicrobial agents or preservativesinclude, but are not limited to, phenols, cresols, mercurials, benzylalcohol, chlorobutanol, methyl and propyl p-hydroxybenzoates,thimerosal, benzalkonium chloride (e.g., benzethonium chloride), methyl-and propyl-parabens, and sorbic acid. Suitable isotonic agents include,but are not limited to, sodium chloride, glycerin, and dextrose.Suitable buffering agents include, but are not limited to, phosphate andcitrate. Suitable antioxidants are those as described herein, includingbisulfite and sodium metabisulfite. Suitable local anesthetics include,but are not limited to, procaine hydrochloride. Suitable suspending anddispersing agents are those as described herein, including sodiumcarboxymethylcelluose, hydroxypropyl methylcellulose, andpolyvinylpyrrolidone. Suitable emulsifying agents include thosedescribed herein, including polyoxyethylene sorbitan monolaurate,polyoxyethylene sorbitan monooleate 80, and triethanolamine oleate.Suitable sequestering or chelating agents include, but are not limitedto EDTA. Suitable pH adjusting agents include, but are not limited to,sodium hydroxide, hydrochloric acid, citric acid, and lactic acid.Suitable complexing agents include, but are not limited to,cyclodextrins, including α-cyclodextrin, β-cyclodextrin,hydroxypropyl-β-cyclodextrin, sulfobutylether-β-cyclodextrin, andsulfobutylether 7-β-cyclodextrin (CAPTISOL®, CyDex, Lenexa, Kans.).

In one embodiment, the pharmaceutical compositions provided herein maybe formulated for single or multiple dosage administration. The singledosage formulations are packaged in an ampoule, a vial, or a syringe.The multiple dosage parenteral formulations may contain an antimicrobialagent at bacteriostatic or fungistatic concentrations. All parenteralformulations must be sterile, as known and practiced in the art.

In one embodiment, the pharmaceutical compositions are provided asready-to-use sterile solutions. In another embodiment, thepharmaceutical compositions are provided as sterile dry solubleproducts, including lyophilized powders and hypodermic tablets, to bereconstituted with a vehicle prior to use. In yet another embodiment,the pharmaceutical compositions are provided as ready-to-use sterilesuspensions. In yet another embodiment, the pharmaceutical compositionsare provided as sterile dry insoluble products to be reconstituted witha vehicle prior to use. In still another embodiment, the pharmaceuticalcompositions are provided as ready-to-use sterile emulsions.

In one embodiment, the pharmaceutical compositions provided herein maybe formulated as immediate or modified release dosage forms, includingdelayed-, sustained, pulsed-, controlled, targeted-, andprogrammed-release forms.

In one embodiment, the pharmaceutical compositions may be formulated asa suspension, solid, semi-solid, or thixotropic liquid, foradministration as an implanted depot. In one embodiment, thepharmaceutical compositions provided herein are dispersed in a solidinner matrix, which is surrounded by an outer polymeric membrane that isinsoluble in body fluids but allows the active ingredient in thepharmaceutical compositions diffuse through.

In one embodiment, suitable inner matrixes includepolymethylmethacrylate, polybutyl-methacrylate, plasticized orunplasticized polyvinylchloride, plasticized nylon, plasticizedpolyethylene terephthalate, natural rubber, polyisoprene,polyisobutylene, polybutadiene, polyethylene, ethylene-vinyl acetatecopolymers, silicone rubbers, polydimethylsiloxanes, silicone carbonatecopolymers, hydrophilic polymers, such as hydrogels of esters of acrylicand methacrylic acid, collagen, cross-linked polyvinyl alcohol, andcross-linked partially hydrolyzed polyvinyl acetate.

In one embodiment, suitable outer polymeric membranes includepolyethylene, polypropylene, ethylene/propylene copolymers,ethylene/ethyl acrylate copolymers, ethylene/vinyl acetate copolymers,silicone rubbers, polydimethyl siloxanes, neoprene rubber, chlorinatedpolyethylene, polyvinylchloride, vinyl chloride copolymers with vinylacetate, vinylidene chloride, ethylene and propylene, ionomerpolyethylene terephthalate, butyl rubber epichlorohydrin rubbers,ethylene/vinyl alcohol copolymer, ethylene/vinyloxyethanol copolymer,and ethylene/vinyl acetate/vinyl alcohol terpolymer.

In specific embodiments, provided herein is a pharmaceutical compositionprepared for parenteral administration (e.g., IV or SC). In oneembodiment, the composition comprises 5-azacytidine as a lyophilizedpowder. In one embodiment, the composition comprises 5-azacytidine andmannitol as a lyophilized powder. In one embodiment, the amount of5-azacytidine in the composition is about 100 mg. In one embodiment, theweight ratio of 5-azacytidine to mannitol is about 1:1. In oneembodiment, the lyophilized powder comprising 5-azacytidine isreconstituted with sterile water for IV or SC administration. In oneembodiment, the dose for parenteral administration is about 75 mg/m². Inone embodiment, the dose for parenteral administration is from about 75mg/m² to about 100 mg/m². In specific embodiments, the composition isadministered daily for 7 days at a dose of about 75 mg/m² to about 100mg/m². In specific embodiments, the composition is administered dailyfor 7 days at a dose of about 75 mg/m² to about 100 mg/m², and the cycleis repeated every 4 weeks. In one embodiment, the compositions isadministered for at least 4 to 6 cycles.

In some embodiments, azacitidine is administered at about 20-200 mg/kgper day (including for example 50 mg/kg, 80 mg/kg, 100 mg/kg, 120 mg/kg,140 mg/kg, 180 mg/kg).

In some embodiments, decitabine is administered at about 0.75-4 mg/kgper day (including for example 1.0 mg/kg, 1.5 mg/kg, 2.00 mg/kg, 2.5mg/kg, 3.0 mg/kg, 3.5 mg/kg).

In some embodiments, azacitidine or decitabine is administered at about10-200 mg/m² (including for example about 50-100 mg/m² or for exampleabout 75 mg/m²).

6. Topical Dosage Forms

In yet another embodiment, the pharmaceutical compositions providedherein may be administered rectally, urethrally, vaginally, orperivaginally in the forms of suppositories, pessaries, bougies,poultices or cataplasm, pastes, powders, dressings, creams, plasters,contraceptives, ointments, solutions, emulsions, suspensions, tampons,gels, foams, sprays, or enemas. These dosage forms can be manufacturedusing conventional processes as described in, e.g., Remington, TheScience and Practice of Pharmacy, supra.

In one embodiment, rectal, urethral, and vaginal suppositories are solidbodies for insertion into body orifices, which are solid at ordinarytemperatures but melt or soften at body temperature to release theactive ingredient(s) inside the orifices. Pharmaceutically acceptablecarriers utilized in rectal and vaginal suppositories include bases orvehicles, such as stiffening agents, which produce a melting point inthe proximity of body temperature, when formulated with thepharmaceutical compositions provided herein; and antioxidants asdescribed herein, including bisulfite and sodium metabisulfite. Suitablevehicles include, but are not limited to, cocoa butter (theobroma oil),glycerin-gelatin, carbowax (polyoxyethylene glycol), spermaceti,paraffin, white and yellow wax, and appropriate mixtures of mono-, di-and triglycerides of fatty acids, hydrogels, such as polyvinyl alcohol,hydroxyethyl methacrylate, polyacrylic acid; glycerinated gelatin.Combinations of the various vehicles may be used. Rectal and vaginalsuppositories may be prepared by the compressed method or molding. Thetypical weight of a rectal and vaginal suppository is about 2 to about 3g.

In one embodiment, the pharmaceutical compositions provided herein maybe administered intranasally or by inhalation to the respiratory tract.The pharmaceutical compositions may be provided in the form of anaerosol or solution for delivery using a pressurized container, pump,spray, atomizer, such as an atomizer using electrohydrodynamics toproduce a fine mist, or nebulizer, alone or in combination with asuitable propellant, such as 1,1,1,2-tetrafluoroethane or1,1,1,2,3,3,3-heptafluoropropane. The pharmaceutical compositions mayalso be provided as a dry powder for insufflation, alone or incombination with an inert carrier such as lactose or phospholipids; andnasal drops. For intranasal use, the powder may comprise a bioadhesiveagent, including chitosan or cyclodextrin.

In one embodiment, solutions or suspensions for use in a pressurizedcontainer, pump, spray, atomizer, or nebulizer may be formulated tocontain ethanol, aqueous ethanol, or a suitable alternative agent fordispersing, solubilizing, or extending release of the active ingredientprovided herein, a propellant as solvent; and/or a surfactant, such assorbitan trioleate, oleic acid, or an oligolactic acid.

In one embodiment, the pharmaceutical compositions provided herein maybe micronized to a size suitable for delivery by inhalation, such asabout 50 micrometers or less, or about 10 micrometers or less. Particlesof such sizes may be prepared using a comminuting method known to thoseskilled in the art, such as spiral jet milling, fluid bed jet milling,supercritical fluid processing to form nanoparticles, high pressurehomogenization, or spray drying.

In one embodiment, capsules, blisters and cartridges for use in aninhaler or insufflator may be formulated to contain a powder mix of thepharmaceutical compositions provided herein; a suitable powder base,such as lactose or starch; and a performance modifier, such asL-leucine, mannitol, or magnesium stearate. The lactose may be anhydrousor in the form of the monohydrate. Other suitable excipients or carriersinclude dextran, glucose, maltose, sorbitol, xylitol, fructose, sucrose,and trehalose. The pharmaceutical compositions provided herein forinhaled/intranasal administration may further comprise a suitableflavor, such as menthol and levomenthol, or sweeteners, such assaccharin or saccharin sodium.

In one embodiment, the pharmaceutical compositions provided herein fortopical administration may be formulated to be immediate release ormodified release, including delayed-, sustained-, pulsed-, controlled-,targeted, and programmed release.

7. Additional Therapeutic Agents

In some embodiments, provided herein is a pharmaceutical compositioncomprising one, two, three, or more other pharmacologically activesubstances (also termed herein “additional therapeutic agents,” “secondactive agents,” or the like) (e.g., other than cytidine analog). In someembodiments, the cytidine analog formulations provided herein furthercomprise one, two, three, or more other pharmacologically activesubstances (also termed herein “additional therapeutic agents,” “secondactive agents,” or the like). In other embodiments, the cytidine analogformulations provided herein is co-administered with one, two, three, ormore other pharmacologically active substances. In particularembodiments, the oral formulations provided herein comprise theadditional therapeutic agent(s) in a therapeutically effective amount.In particular embodiments, the cytidine analog (e.g., 5-azacytidine oranother cytidine analog provided herein) and the additional therapeuticagent(s) are co-formulated together in the same dosage form usingmethods of co-formulating active pharmaceutical ingredients, includingmethods disclosed herein and methods known in the art. In otherembodiments, the cytidine analog (e.g., 5-azacytidine or anothercytidine analog provided herein) and the additional therapeutic agent(s)are co-administered in separate dosage forms. It is believed thatcertain combinations work synergistically in the treatment of particulardiseases or disorders, including, e.g., types of cancer and certaindiseases and conditions associated with, or characterized by, undesiredangiogenesis or abnormal cell proliferation, for example, solid tumors.Cytidine analog dosage forms provided herein can also work to alleviateadverse effects associated with certain second active agents, and somesecond active agents can be used to alleviate adverse effects associatedwith cytidine analog dosage forms provided herein. In certainembodiments, the formulations of cytidine analogs provided herein areco-administered with one or more therapeutic agents to provide aresensitization effect in subjects in need thereof. Additionaltherapeutic agents can be, e.g., large molecules (e.g., proteins) orsmall molecules (e.g., synthetic inorganic, organometallic, or organicmolecules).

Examples of particular additional therapeutic agents useful in thecompositions and methods disclosed herein include, but are not limitedto, e.g., cytotoxic agents, anti-metabolites, antifolates, HDACinhibitors (e.g., entinostat, also known as SNDX-275 or MS-275; orvorinostat, also known as suberoylanilide hydroxamic acid (SAHA) orN-hydroxy-N-phenyl-octanediamide), DNA intercalating agents, DNAcross-linking agents, DNA alkylating agents, DNA cleaving agents,topoisomerase inhibitors, CDK inhibitors, JAK inhibitors,anti-angiogenic agents, Bcr-Abl inhibitors, HER2 inhibitors, EGFRinhibitors, VEGFR inhibitors, PDGFR inhibitors, HGFR inhibitors, IGFRinhibitors, c-Kit inhibitors, Ras pathway inhibitors, PI3K inhibitors,multi-targeted kinase inhibitors, mTOR inhibitors, anti-estrogens,anti-androgens, aromatase inhibitors, somatostatin analogs, ERmodulators, anti-tubulin agents, vinca alkaloids, taxanes, HSPinhibitors, Smoothened antagonists, telomerase inhibitors, COX-2inhibitors, anti-metastatic agents, immunosuppressants, biologics suchas antibodies, and hormonal therapies. In particular embodiments, theco-administered therapeutic agent is an immunomodulatory compound, e.g.,thalidomide, lenalidomide, or pomalidomide. In particular embodiments,the co-administered therapeutic agent is carboplatin. In particularembodiments, the co-administered therapeutic agent is paclitaxel (e.g.,Abraxane®). See, e.g., U.S. Pat. Nos. 7,758,891, 7,771,751, 7,820,788,7,923,536, 8,034,375; U.S. Patent Publication No. 2010/0048499; see alsoU.S. Pat. Nos. 5,916,596; 6,506,405; 6,749,868, and 6,537,579, and U.S.Pat. Pub. Nos. 2007/0082838; all of which are incorporated herein byreference in their entireties. Other references include PCT ApplicationPublication Nos. WO08/057562, WO09/126938, WO09/126401, WO09/126175,incorporated herein by reference. In one embodiment, the co-administeredagent may be dosed, e.g., orally or by injection.

In one embodiment, the additional therapeutic agent is a compositioncomprising nanoparticles comprising a taxane (such as paclitaxel) and acarrier protein. In some embodiments, the nanoparticle compositioncomprises nanoparticles comprising paclitaxel and an albumin. In someembodiments, the nanoparticles in the composition described herein havean average diameter of no greater than about 200 nm, including forexample no greater than about any one of 190, 180, 170, 160, 150, 140,130, 120, 110, 100, 90, 80, 70, or 60 nm. In some embodiments, at leastabout 50% (for example at least about any one of 60%, 70%, 80%, 90%,95%, or 99%) of all the nanoparticles in the composition have a diameterof no greater than about 200 nm, including for example no greater thanabout any one of 190, 180, 170, 160, 150, 140, 130, 120, 110, 100, 90,80, 70, or 60 nm. In some embodiments, at least about 50% (for exampleat least any one of 60%, 70%, 80%, 90%, 95%, or 99%) of all thenanoparticles in the composition fall within the range of about 20 toabout 400, including for example about 20 to about 200 nm, about 30 toabout 180 nm, and any one of about 40 to about 150, about 50 to about120, and about 60 to about 100 nm. In some embodiments, the carrierprotein has sulfhydral groups that can form disulfide bonds. In someembodiments, at least about 5% (including for example at least about anyone of 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%) of thecarrier protein in the nanoparticle portion of the composition arecrosslinked (for example crosslinked through one or more disulfidebonds).

In some embodiments, the nanoparticles comprise the taxane (such aspaclitaxel) coated with a carrier protein, such as albumin (e.g., humanserum albumin). In some embodiments, the composition comprises taxane inboth nanoparticle and non-nanoparticle form, wherein at least about anyone of 50%, 60%, 70%, 80%, 90%, 95%, or 99% of the taxane in thecomposition are in nanoparticle form. In some embodiments, the taxane inthe nanoparticles constitutes more than about any one of 50%, 60%, 70%,80%, 90%, 95%, or 99% of the nanoparticles by weight. In someembodiments, the nanoparticles comprise a core of taxane that issubstantially free of polymeric materials (such as polymeric matrix).

In some embodiments, the nanoparticle composition is substantially free(such as free) of surfactants (such as Cremophor®, Tween 80, or otherorganic solvents used for the administration of taxanes). In someembodiments, the nanoparticle composition contains less than about anyone of 20%, 15%, 10%, 7.5%, 5%, 2.5%, or 1% organic solvent. In someembodiments, the weight ratio of carrier protein (such as albumin) andtaxane in the nanoparticle composition is about 18:1 or less, such asabout 15:1 or less, for example about 9:1 or less. In some embodiments,the weight ratio of carrier protein (such as albumin) and taxane in thecomposition falls within the range of any one of about 1:1 to about18:1, about 2:1 to about 15:1, about 3:1 to about 13:1, about 4:1 toabout 12:1, about 5:1 to about 10:1, about 9:1. In some embodiments, theweight ratio of carrier protein and taxane in the nanoparticle portionof the composition is about any one of 1:2, 1:3, 1:4, 1:5, 1:9, 1:10,1:15, or less.

In some embodiments, the particle composition comprises one or more ofthe above characteristics. In some embodiments, the nanoparticlecomposition is Abraxane®. Nanoparticle compositions comprising othertaxanes (such as docetaxel and ortataxel) may also comprise one or moreof the above characteristics.

In some embodiments, Abraxane® is intravenously administering at a doseof about 80 to about 200 mg/m² (such as about 100 mg/m²). In someembodiments, Abraxane® is administered weekly. In some embodiments,Abraxane® is administered once every two weeks. In some embodiments,Abraxane® is administered once every three weeks. In some embodiments,Abraxane® is administered as part of a cyclic treatment regimen (e.g.,in cycles). In some embodiments, Abraxane® is administered on Days 1 and8 of a 21-day cycle. In some embodiments, Abraxane® is administered onDays 8 and 15 of a 21-day cycle.

In some embodiment, carboplatin is intravenously administering at thedose of about AUC 2 to AUC 6 (such as AUC 2, AUC 4, AUC 6). In someembodiments, carboplatin is administered weekly. In some embodiments,carboplatin is administered once every two weeks. In some embodiments,carboplatin is administered once every three weeks. In some embodiments,carboplatin is administered as part of a cyclic treatment regimen (e.g.,in cycles).

Other examples of additional therapeutic agents include, but are notlimited to, hematopoietic growth factor, a cytokine, an anti-canceragent, granulocyte colony-stimulating factor (G-CSF),granulocyte-macrophage colony-stimulating factor (GM-CSF),erythropoietin (EPO), interleukin (IL), interferon (IFN), oblimersen,melphalan, topotecan, pentoxifylline, taxotere, irinotecan,ciprofloxacin, doxorubicin, vincristine, dacarbazine, Ara-C,vinorelbine, prednisone, cyclophosphamide, bortezomib, arsenic trioxide.Such additional therapeutic agents are particularly useful in methodsand compositions disclosed herein including, but not limited to, thoserelating to treatment of multiple myeloma.

Other examples of additional therapeutic agents include, but are notlimited to, an antibody (e.g., rituximab, anti-CD33), hematopoieticgrowth factor, cytokine, anti-cancer agent, antibiotic, cox-2 inhibitor,immunomodulatory agent, immunosuppressive agent, corticosteroid, or apharmacologically active mutant or derivative thereof. See, e.g., S.Nand et al., Leukemia and Lymphoma, 2008, 49(11):2141-47 (describing aPhase II study involving the administration of a combination ofhydroxyurea, azacitidine and low dose gemtuzumab ozogamicin to elderlypatients with AML and high-risk MDS, and concluding that thiscombination appears to be a safe and effective regimen in the treatmentof AML and high risk MDS in this group of patients). Such additionaltherapeutic agents are particularly useful in methods and compositionsdisclosed herein including, but not limited to, those relating totreatment of the diseases and disorders disclosed herein.

Examples of large molecule active agents include, but are not limitedto, hematopoietic growth factors, cytokines, and monoclonal andpolyclonal antibodies. Typical large molecule active agents arebiological molecules, such as naturally occurring or artificially madeproteins. Proteins that are particularly useful include proteins thatstimulate the survival and/or proliferation of hematopoietic precursorcells and immunologically active poietic cells in vitro or in vivo.Others stimulate the division and differentiation of committed erythroidprogenitors in cells in vitro or in vivo. Particular proteins include,but are not limited to: interleukins, such as IL-2 (includingrecombinant IL-II (“rIL2”) and canarypox IL-2), IL-10, IL-12, and IL-18;interferons, such as interferon alfa-2a, interferon alfa-2b, interferonalfa-n1, interferon alfa-n3, interferon beta-I a, and interferon gamma-Ib; GM-CF and GM-CSF; and EPO.

Particular proteins that can be used in the methods and compositionsprovided herein include, but are not limited to: filgrastim, which issold in the United States under the trade name Neupogen® (Amgen,Thousand Oaks, Calif.); sargramostim, which is sold in the United Statesunder the trade name Leukine® (Immunex, Seattle, Wash.); and recombinantEPO, which is sold in the United States under the trade name Epogen®(Amgen, Thousand Oaks, Calif.).

Recombinant and mutated forms of GM-CSF can be prepared as described inU.S. Pat. Nos. 5,391,485; 5,393,870; and 5,229,496; all of which areincorporated herein by reference. Recombinant and mutated forms of G-CSFcan be prepared as described in U.S. Pat. Nos. 4,810,643; 4,999,291;5,528,823; and 5,580,755; all of which are incorporated herein byreference.

Embodiments herein encompass the use of native, naturally occurring, andrecombinant proteins. Particular embodiments encompass mutants andderivatives (e.g., modified forms) of naturally occurring proteins thatexhibit, in vivo, at least some of the pharmacological activity of theproteins upon which they are based. Examples of mutants include, but arenot limited to, proteins that have one or more amino acid residues thatdiffer from the corresponding residues in the naturally occurring formsof the proteins. Also encompassed by the term “mutants” are proteinsthat lack carbohydrate moieties normally present in their naturallyoccurring forms (e.g., nonglycosylated forms). Examples of derivativesinclude, but are not limited to, pegylated derivatives and fusionproteins, such as proteins formed by fusing IgG1 or IgG3 to the proteinor active portion of the protein of interest. See, e.g., Penichet, M. L.and Morrison, S. L., J. Immunol. Methods 248:91-101 (2001).

Antibodies that can be used in combination with oral formulationsdisclosed herein include monoclonal and polyclonal antibodies. Examplesof antibodies include, but are not limited to, trastuzumab (Herceptin®),rituximab (Rituxan®), bevacizumab (Avastin™), pertuzumab (Omnitarg™),tositumomab (Bexxar®), edrecolomab (Panorex®), and G250. Oralformulations disclosed herein can also comprise, be combined with, orused in combination with anti-TNF-α antibodies.

Large molecule active agents may be administered in the form ofanti-cancer vaccines. For example, vaccines that secrete, or cause thesecretion of, cytokines such as IL-2, G-CSF, and GM-CSF can be used inthe methods, pharmaceutical compositions, and kits provided herein. See,e.g., Emens, L. A., et al., Curr. Opinion Mol. Ther. 3(1):77-84 (2001).

In one embodiment, the additional therapeutic agent (e.g.,large-molecule compound or small-molecule compound) reduces, eliminates,or prevents an adverse effect associated with the administration (e.g.,oral administration) of a cytidine analog provided herein. Depending onthe particular cytidine analog and the disease or disorder begintreated, adverse effects can include, but are not limited to, anemia,neutropenia, febrile neutropenia, thrombocytopenia, hepatotoxicity(e.g., including, but not limited to, hepatoxicity in patients withpreexisting hepatic impairment), elevated serum creatinine, renalfailure, renal tubular acidosis, hypokalemia, hepatic coma, nausea,vomiting, dyspepsia, abdominal pain, pyrexia, leukopenia, diarrhea,constipation, ecchymosis, petechiae, rigors, weakness, pneumonia,anxiety, insomnia, lethargy, and decrease in weight, among others knownin the art to be associated with particular cytidine analogs.

Like some large molecules, many small-molecule compounds are believed tobe capable of providing a synergistic effect when administered with(e.g., before, after or simultaneously) a cytidine analog oralformulation disclosed herein. Examples of small molecule second activeagents include, but are not limited to, anti-cancer agents, antibiotics,immunosuppressive agents, and steroids.

Examples of anti-cancer agents include, but are not limited to:acivicin; aclarubicin; acodazole hydrochloride; acronine; adozelesin;aldesleukin; altretamine; ambomycin; ametantrone acetate; amsacrine;anastrozole; anthramycin; asparaginase; asperlin; azacitidine; azetepa;azotomycin; batimastat; benzodepa; bicalutamide; bisantrenehydrochloride; bisnafide dimesylate; bizelesin; bleomycin sulfate;brequinar sodium; bropirimine; busulfan; cactinomycin; calusterone;caracemide; carbetimer; carboplatin; carmustine; carubicinhydrochloride; carzelesin; cedefingol; celecoxib (COX-2 inhibitor);chlorambucil; cirolemycin; cisplatin; cladribine; crisnatol mesylate;cyclophosphamide; cytarabine; dacarbazine; dactinomycin; daunorubicinhydrochloride; decitabine; dexormaplatin; dezaguanine; dezaguaninemesylate; diaziquone; docetaxel; doxorubicin; doxorubicin hydrochloride;droloxifene; droloxifene citrate; dromostanolone propionate; duazomycin;edatrexate; eflornithine hydrochloride; elsamitrucin; enloplatin;enpromate; epipropidine; epirubicin hydrochloride; erbulozole;esorubicin hydrochloride; estramustine; estramustine phosphate sodium;etanidazole; etoposide; etoposide phosphate; etoprine; fadrozolehydrochloride; fazarabine; fenretinide; floxuridine; fludarabinephosphate; fluorouracil; flurocitabine; fosquidone; fostriecin sodium;gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicinhydrochloride; ifosfamide; ilmofosine; iproplatin; irinotecan;irinotecan hydrochloride; lanreotide acetate; letrozole; leuprolideacetate; liarozole hydrochloride; lometrexol sodium; lomustine;losoxantrone hydrochloride; masoprocol; maytansine; mechlorethaminehydrochloride; megestrol acetate; melengestrol acetate; melphalan;menogaril; mercaptopurine; methotrexate; methotrexate sodium; metoprine;meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin; mitomalcin;mitomycin; mitosper; mitotane; mitoxantrone hydrochloride; mycophenolicacid; nocodazole; nogalamycin; ormaplatin; oxisuran; paclitaxel;pegaspargase; peliomycin; pentamustine; peplomycin sulfate;perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride;plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine;procarbazine hydrochloride; puromycin; puromycin hydrochloride;pyrazofurin; riboprine; safingol; safingol hydrochloride; semustine;simtrazene; sparfosate sodium; sparsomycin; spirogermaniumhydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin;sulofenur; talisomycin; tecogalan sodium; taxotere; tegafur;teloxantrone hydrochloride; temoporfin; teniposide; teroxirone;testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin;tirapazamine; toremifene citrate; trestolone acetate; triciribinephosphate; trimetrexate; trimetrexate glucuronate; triptorelin;tubulozole hydrochloride; uracil mustard; uredepa; vapreotide;verteporfin; vinblastine sulfate; vincristine sulfate; vindesine;vindesine sulfate; vinepidine sulfate; vinglycinate sulfate;vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate;vinzolidine sulfate; vorozole; zeniplatin; zinostatin; and zorubicinhydrochloride.

Other anti-cancer drugs include, but are not limited to: 20-epi-1,25dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin;acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TK antagonists;altretamine; ambamustine; amidox; amifostine; aminolevulinic acid;amrubicin; amsacrine; anagrelide; anastrozole; andrographolide;angiogenesis inhibitors; antagonist D; antagonist G; antarelix;anti-dorsalizing morphogenetic protein-1; antiandrogen, prostaticcarcinoma; antiestrogen; antineoplaston; antisense oligonucleotides;aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators;apurinic acid; ara-CDP-DL-PTBA; arginine deaminase; asulacrine;atamestane; atrimustine; axinastatin 1; axinastatin 2; axinastatin 3;azasetron; azatoxin; azatyrosine; baccatin III derivatives; balanol;batimastat; BCR/ABL antagonists; benzochlorins; benzoylstaurosporine;beta lactam derivatives; beta-alethine; betaclamycin B; betulinic acid;bFGF inhibitor; bicalutamide; bisantrene; bisaziridinylspermine;bisnafide; bistratene A; bizelesin; breflate; bropirimine; budotitane;buthionine sulfoximine; calcipotriol; calphostin C; camptothecinderivatives; capecitabine; carboxamide-amino-triazole;carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived inhibitor;carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropinB; cetrorelix; chlorins; chloroquinoxaline sulfonamide; cicaprost;cis-porphyrin; cladribine; clomifene analogues; clotrimazole;collismycin A; collismycin B; combretastatin A4; combretastatinanalogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8;cryptophycin A derivatives; curacin A; cyclopentanthraquinones;cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor;cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin;dexamethasone; dexifosfamide; dexrazoxane; dexverapamil; diaziquone;didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine;dihydrotaxol, 9-; dioxamycin; diphenyl spiromustine; docetaxel;docosanol; dolasetron; doxifluridine; doxorubicin; droloxifene;dronabinol; duocarmycin SA; ebselen; ecomustine; edelfosine;edrecolomab; eflornithine; elemene; emitefur; epirubicin; epristeride;estramustine analogue; estrogen agonists; estrogen antagonists;etanidazole; etoposide phosphate; exemestane; fadrozole; fazarabine;fenretinide; filgrastim; finasteride; flavopiridol; flezelastine;fluasterone; fludarabine; fluorodaunorunicin hydrochloride; forfenimex;formestane; fostriecin; fotemustine; gadolinium texaphyrin; galliumnitrate; galocitabine; ganirelix; gelatinase inhibitors; gemcitabine;glutathione inhibitors; hepsulfam; heregulin; hexamethylenebisacetamide; hypericin; ibandronic acid; idarubicin; idoxifene;idramantone; ilmofosine; ilomastat; imatinib (e.g., Gleevec®),imiquimod; immunostimulant peptides; insulin-like growth factor-1receptor inhibitor; interferon agonists; interferons; interleukins;iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact; irsogladine;isobengazole; isohomohalicondrin B; itasetron; jasplakinolide;kahalalide F; lamellarin-N triacetate; lanreotide; leinamycin;lenograstim; lentinan sulfate; leptolstatin; letrozole; leukemiainhibiting factor; leukocyte alpha interferon;leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;linear polyamine analogue; lipophilic disaccharide peptide; lipophilicplatinum compounds; lissoclinamide 7; lobaplatin; lombricine;lometrexol; lonidamine; losoxantrone; loxoribine; lurtotecan; lutetiumtexaphyrin; lysofylline; lytic peptides; maitansine; mannostatin A;marimastat; masoprocol; maspin; matrilysin inhibitors; matrixmetalloproteinase inhibitors; menogaril; merbarone; meterelin;methioninase; metoclopramide; MIF inhibitor; mifepristone; miltefosine;mirimostim; mitoguazone; mitolactol; mitomycin analogues; mitonafide;mitotoxin fibroblast growth factor-saporin; mitoxantrone; mofarotene;molgramostim; Erbitux, human chorionic gonadotrophin; monophosphoryllipid A+myobacterium cell wall sk; mopidamol; mustard anticancer agent;mycaperoxide B; mycobacterial cell wall extract; myriaporone;N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip;naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin;nemorubicin; neridronic acid; nilutamide; nisamycin; nitric oxidemodulators; nitroxide antioxidant; nitrullyn; oblimersen (Genasense®);O⁶-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone;ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin;osaterone; oxaliplatin; oxaunomycin; paclitaxel; paclitaxel analogues;paclitaxel derivatives; palauamine; palmitoylrhizoxin; pamidronic acid;panaxytriol; panomifene; parabactin; pazelliptine; pegaspargase;peldesine; pentosan polysulfate sodium; pentostatin; pentrozole;perflubron; perfosfamide; perillyl alcohol; phenazinomycin;phenylacetate; phosphatase inhibitors; picibanil; pilocarpinehydrochloride; pirarubicin; piritrexim; placetin A; placetin B;plasminogen activator inhibitor; platinum complex; platinum compounds;platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;propyl bis-acridone; prostaglandin J2; proteasome inhibitors; proteinA-based immune modulator; protein kinase C inhibitor; protein kinase Cinhibitors, microalgal; protein tyrosine phosphatase inhibitors; purinenucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists;raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors;ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re186 etidronate; rhizoxin; ribozymes; RII retinamide; rohitukine;romurtide; roquinimex; rubiginone Bl; ruboxyl; safingol; saintopin;SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics; semustine;senescence derived inhibitor 1; sense oligonucleotides; signaltransduction inhibitors; sizofiran; sobuzoxane; sodium borocaptate;sodium phenylacetate; solverol; somatomedin binding protein; sonermin;sparfosic acid; spicamycin D; spiromustine; splenopentin; spongistatin1; squalamine; stipiamide; stromelysin inhibitors; sulfinosine;superactive vasoactive intestinal peptide antagonist; suradista;suramin; swainsonine; tallimustine; tamoxifen methiodide; tauromustine;tazarotene; tecogalan sodium; tegafur; tellurapyrylium; telomeraseinhibitors; temoporfin; teniposide; tetrachlorodecaoxide; tetrazomine;thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroidstimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocenebichloride; topsentin; toremifene; translation inhibitors; tretinoin;triacetyluridine; triciribine; trimetrexate; triptorelin; tropisetron;turosteride; tyrosine kinase inhibitors; tyrphostins; UBC inhibitors;ubenimex; urogenital sinus-derived growth inhibitory factor; urokinasereceptor antagonists; vapreotide; variolin B; velaresol; veramine;verdins; verteporfin; vinorelbine; vinxaltine; vitaxin; vorozole;zanoterone; zeniplatin; zilascorb; and zinostatin stimalamer.

Specific additional therapeutic agents include, but are not limited to,oblimersen (Genasense®), remicade, docetaxel, celecoxib, melphalan,dexamethasone (Decadron®), steroids, gemcitabine, cisplatinum,temozolomide, etoposide, cyclophosphamide, temodar, carboplatin,procarbazine, gliadel, tamoxifen, topotecan, methotrexate, Arisa®,taxol, taxotere, fluorouracil, leucovorin, irinotecan, xeloda, CPT-11,interferon alpha, pegylated interferon alpha (e.g., PEG INTRON-A),capecitabine, cisplatin, thiotepa, fludarabine, carboplatin, liposomaldaunorubicin, cytarabine, doxetaxol, pacilitaxel, vinblastine, IL-2,GM-CSF, dacarbazine, vinorelbine, zoledronic acid, palmitronate, biaxin,busulphan, prednisone, bisphosphonate, arsenic trioxide, vincristine,doxorubicin (Doxil), paclitaxel, ganciclovir, adriamycin, estramustinesodium phosphate (Emcyt®), sulindac, and etoposide.

8. Kits

In one embodiment, active ingredients provided herein are notadministered to a patient at the same time or by the same route ofadministration. Provided herein are kits which can simplify theadministration of appropriate amounts of active ingredients.

In one embodiment, a kit comprises a dosage form of a compound providedherein. Kits can further comprise one or more second active ingredientsas described herein, or a pharmacologically active mutant or derivativethereof, or a combination thereof.

In other embodiments, kits can further comprise devices that are used toadminister the active ingredients. Examples of such devices include, butare not limited to, syringes, drip bags, patches, and inhalers.

In one embodiment, kits can further comprise cells or blood fortransplantation as well as pharmaceutically acceptable vehicles that canbe used to administer one or more active ingredients. For example, if anactive ingredient is provided in a solid form that must be reconstitutedfor parenteral administration, the kit can comprise a sealed containerof a suitable vehicle in which the active ingredient can be dissolved toform a particulate-free sterile solution that is suitable for parenteraladministration. Examples of pharmaceutically acceptable vehiclesinclude, but are not limited to: Water for Injection USP; aqueousvehicles such as, but not limited to, Sodium Chloride Injection,Ringer's Injection, Dextrose Injection, Dextrose and Sodium ChlorideInjection, and Lactated Ringer's Injection; water-miscible vehicles suchas, but not limited to, ethyl alcohol, polyethylene glycol, andpolypropylene glycol; and non-aqueous vehicles such as, but not limitedto, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate,isopropyl myristate, and benzyl benzoate.

D. METHODS OF USE

In one embodiment, provided herein are methods for treating, preventingor managing cancer by administering a cytidine analog, or apharmaceutically acceptable salt, solvate, or hydrate thereof, to asubject having cancer. In one embodiment, the methods comprise treatingcancer with a cytidine analog, or a pharmaceutically acceptable salt,solvate, or hydrate thereof. In one embodiment, the methods comprisepreventing cancer with a cytidine analog, or a pharmaceuticallyacceptable salt, solvate, or hydrate thereof. In one embodiment, themethods comprise managing cancer with a cytidine analog, or apharmaceutically acceptable salt, solvate, or hydrate thereof. Incertain embodiments, the cytidine analog is 5-azacytidine. In certainembodiments, the cytidine analog is decitabine. In certain embodiments,the methods comprise co-administering one or more additional activeagents (e.g., an anti-cancer agent provided herein). In certainembodiments, the subject is a mammal. In certain embodiments, thesubject is a human. In particular embodiments, the cancer is a solidtumor (e.g., a relapsed or refractory solid tumor).

In one embodiment, provided herein is use of a cytidine analog (e.g.,5-azacytidine or another cytidine analog provided herein), or apharmaceutically acceptable salt, solvate, or hydrate thereof, in themanufacture of a medicament for the treatment, prevention, and/ormanagement of cancer (e.g., a relapsed or refractory solid tumor).

In one embodiment, provided herein is a cytidine analog (e.g.,5-azacytidine or another cytidine analog provided herein), or apharmaceutically acceptable salt, solvate, or hydrate thereof, for usein the treatment, prevention, and/or management of cancer (e.g., arelapsed or refractory solid tumor).

In one embodiment, provided herein are methods of treating, preventing,or managing certain types of cancer, including but not limited to, asolid tumor or a blood-borne tumor; a refractory cancer or a relapsedcancer; or a refractory solid tumor or a relapsed solid tumor. In oneembodiment, provided herein are methods of treating, preventing, ormanaging certain types of cancer, including but not limited to, cancersof the breast, lung, head and neck, ovary, testicle, prostate,gastrointestinal system, stomach, pancreas, liver, colon, kidney,bladder, brain, skin, or bone. In other embodiments, the cancer is acancer of the blood or the lymph.

In one embodiment, provided herein is a method for treating a subjecthaving a cancer, wherein the method comprises orally administering tothe subject a pharmaceutical composition comprising 5-azacytidine, or apharmaceutically acceptable salt, solvate, or hydrate thereof; and themethod optionally further comprises administering at least oneadditional therapeutic agent.

In one embodiment, provided herein are methods of treating, preventing,or managing gastrointentinal stromal tumor, moderately differentiatedadenocarcinoma, metastatic colorectal adenocarcinoma, sigmoid colonadenocarcinoma, appendicial mucinous adenocarcinoma, metastaticcolorectal cancer, nasopharyngeal carcinoma, cervical carcinoma,cervical adenocarcinoma, non-small cell adenocarcinoma of the lung,colon Kras adenocarcinoma, and colon adenocarcinoma, comprising orallyadministering 5-azacytidine and optionally at least one additionalthereapeutic agent.

In one embodiment, provided herein are methods of treating, preventing,or managing gastrointentinal stromal tumor, moderately differentiatedadenocarcinoma, metastatic colorectal adenocarcinoma, sigmoid colonadenocarcinoma, appendicial mucinous adenocarcinoma, metastaticcolorectal cancer, nasopharyngeal carcinoma, cervical carcinoma,cervical adenocarcinoma, non-small cell adenocarcinoma of the lung,colon Kras adenocarcinoma, and colon adenocarcinoma, comprising orallyadministering 5-azacytidine.

In one embodiment, provided herein are methods of treating, preventing,or managing nasopharyngeal carcinoma, cervical carcinoma, cervicaladenocarcinoma, non-small cell adenocarcinoma of the lung, colon Krasadenocarcinoma, and colon adenocarcinoma, comprising orallyadministering 5-azacytidine.

In one embodiment, provided herein are methods of treating, preventing,or managing adenocarcinoma of colon, breast ductal carcinoma, gallbladder adenocarcinoma, pancreatic adenocarcinoma, pancreaticcancer,metastatic pancreatic adenocarcinoma, endometrial carcinoma,ovarian cancer, metastatic ovarian carcinoma, papillary serousendocervical adenocarcinoma, endometroid type well-differentiatedadenocarcinoma, rectal adenocarcinoma, pancreatic carcinoma, comprisingorally administering 5-azacytidine and optionally at least oneadditional thereapeutic agent.

In one embodiment, provided herein are methods of treating, preventing,or managing adenocarcinoma of colon, breast ductal carcinoma, gallbladder adenocarcinoma, pancreatic adenocarcinoma, pancreatic cancer,metastatic pancreatic adenocarcinoma, endometrial carcinoma, ovariancancer, metastatic ovarian carcinoma, papillary serous endocervicaladenocarcinoma, endometroid type well-differentiated adenocarcinoma,rectal adenocarcinoma, pancreatic carcinoma, and metastatic pancreaticadenocarcinoma, comprising orally administering 5-azacytidine and ABX.

In one embodiment, provided herein are methods of treating, preventing,or managing pancreatic cancer, metastatic pancreatic adenocarcinoma,endometrial carcinoma, ovarian cancer, metastatic ovarian carcinoma,papillary serous endocervical adenocarcinoma, endometroid typewell-differentiated adenocarcinoma, rectal adenocarcinoma, andpancreatic carcinoma, comprising orally administering 5-azacytidine andABX.

In one embodiment, provided herein are methods of treating, preventing,or managing Burkitt's lymphoma, Hodgkin's lymphoma, nasopharyngealcarcinoma, cervical carcinoma, head and neck squamous cell carcinoma,Merkel cell tumors, adenocarcinoma of the colon, pancreatic cancer, gallbladder adenocarcinoma, pancreatic adenocarcinoma, metastatic pancreaticadenocarcinoma, endometrial carcinoma, ovarian cancer, pancreaticcarcinoma, metastatic ovarian carcinoma, gastrointestinal stromal tumor,sigmoid colon adenocarcinoma, metastatic colorectal cancer, breastductal carcinoma, papillary serous endocervical adenocarcinoma,endometroid type well-differentiated adenocarcinoma, rectaladenocarcinoma, moderately differentiated adenocarcinoma, metastaticcolorectal adenocarcinoma, appendicial mucinous adenocarcinoma, cervicaladenocarcinoma, non-small cell adenocarcinoma of the lung, colon Krasadenocarcinoma, and colon adenocarcinoma, comprising orallyadministering 5-azacytidine and optionally at least one additionalthereapeutic agent.

In one embodiment, provided herein are methods of treating, preventing,or managing breast cancer, comprising administering a cytidine analog(e.g., orally) and at least one additional therapeutic agent (e.g.,additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing lung cancer (e.g., NSCLC or SCLC), comprising administeringa cytidine analog (e.g., orally) and at least one additional therapeuticagent (e.g., additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing head and neck cancer, comprising administering a cytidineanalog (e.g., orally) and at least one additional therapeutic agent(e.g., additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing ovarian cancer, comprising administering a cytidine analog(e.g., orally) and at least one additional therapeutic agent (e.g.,additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing testicular cancer, comprising administering a cytidineanalog (e.g., orally) and at least one additional therapeutic agent(e.g., additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing prostate cancer, comprising administering a cytidine analog(e.g., orally) and at least one additional therapeutic agent (e.g.,additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing gastrointestinal cancer, comprising administering a cytidineanalog (e.g., orally) and at least one additional therapeutic agent(e.g., additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing stomach cancer, comprising administering a cytidine analog(e.g., orally) and at least one additional therapeutic agent (e.g.,additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing pancreatic cancer, comprising administering a cytidineanalog (e.g., orally) and at least one additional therapeutic agent(e.g., additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing liver cancer, comprising administering a cytidine analog(e.g., orally) and at least one additional therapeutic agent (e.g.,additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing colorectal cancer, comprising administering a cytidineanalog (e.g., orally) and at least one additional therapeutic agent(e.g., additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing renal cancer, comprising administering a cytidine analog(e.g., orally) and at least one additional therapeutic agent (e.g.,additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing bladder cancer, comprising administering a cytidine analog(e.g., orally) and at least one additional therapeutic agent (e.g.,additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing brain cancer, comprising administering a cytidine analog(e.g., orally) and at least one additional therapeutic agent (e.g.,additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing skin cancer (e.g., melanoma), comprising administering acytidine analog (e.g., orally) and at least one additional therapeuticagent (e.g., additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing bone cancer, comprising administering a cytidine analog(e.g., orally) and at least one additional therapeutic agent (e.g.,additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing blood cancer, comprising administering a cytidine analog(e.g., orally) and at least one additional therapeutic agent (e.g.,additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing leukemia, comprising administering a cytidine analog (e.g.,orally) and at least one additional therapeutic agent (e.g., additionaltherapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing lymphoma, comprising administering a cytidine analog (e.g.,orally) and at least one additional therapeutic agent (e.g., additionaltherapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing multiple myeloma, comprising administering a cytidine analog(e.g., orally) and at least one additional therapeutic agent (e.g.,additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing myelodysplastic syndrome, comprising administering acytidine analog (e.g., orally) and at least one additional therapeuticagent (e.g., additional therapeutic agent described herein).

In one embodiment, provided herein are methods of treating, preventing,or managing cancer in the primary tumor, lymph nodes, or distantmetastasis, by administering a cytidine analog, or a pharmaceuticallyacceptable salt, solvate, or hydrate thereof, to a subject in needthereof. In one embodiment, provided herein are methods of treating,preventing, or managing cancer in the primary tumor, lymph nodes, ordistant metastasis, by administering 5-azacytidine, or apharmaceutically acceptable salt, solvate, or hydrate thereof, to asubject in need thereof.

In one embodiment, provided herein are methods of treating, preventing,or managing cancer in a subject having surgically resectable cancer,locally or regionally advanced cancer, or distant metastatic cancer, byadministering a cytidine analog, or a pharmaceutically acceptable salt,solvate, or hydrate thereof. In one embodiment, provided herein aremethods of treating, preventing, or managing cancer in a subject havingsurgically resectable cancer, locally or regionally advanced cancer, ordistant metastatic cancer, by administering 5-azacytidine, or apharmaceutically acceptable salt, solvate, or hydrate thereof. Inparticular embodiments, provided herein are methods of treatingsurgically resectable cancer, by administering 5-azacytidine to asubject having cancer. In particular embodiments, provided herein aremethods of treating locally or regionally advanced cancer, byadministering 5-azacytidine to a subject having cancer. In particularembodiments, provided herein are methods of treating distant metastaticcancer, by administering 5-azacytidine to a subject having cancer.

In one embodiment, the methods comprise treating, preventing or managingcertain stages of cancer, e.g., Stage 0, Stage I, Stage II, Stage III,and Stage IV, by administering a cytidine analog, or a pharmaceuticallyacceptable salt, solvate, or hydrate thereof, to a subject havingcancer. The staging of cancer may be defined according to methods knownin the art, for example, according to the guidelines provided by theAmerican Joint Committee on Cancer (AJCC). In one embodiment, thestaging of cancer is designated and grouped based on the TNMclassification, i.e., a classification based on the status of primarytumor (e.g., TX, T0, Tis, T1, T2, T3, T4), regional lymph nodes (e.g.,NX, N0, N1, N2, N3), and/or distant metastasis (e.g., MX, M0, M1), in asubject having cancer.

In particular embodiments, methods provided herein compriseadministering a cytidine analog to a subject having a solid tumor thatis surgically resectable. In particular embodiments, the methodsprovided herein comprise administering a cytidine analog to a subjecthaving locally advanced solid tumor. In particular embodiments, methodsprovided herein comprise administering a cytidine analog to a subjecthaving regionally advanced solid tumor. In particular embodiments, themethods provided herein comprise administering a cytidine analog to asubject having a distant metastasis, e.g., at the time of diagnosis.

Particular embodiments provide treating a subject having cancer usingone or more of the methods provided herein, together with surgery.Particular embodiments provide treating a subject having cancer usingone or more of the methods provided herein, together with chemotherapy.Particular embodiments provide treating a subject having cancer usingone or more of the methods provided herein, together with immunotherapy.Particular embodiments provide treating a subject having cancer usingone or more of the methods provided herein, together with targetedtherapy. Particular embodiments provide treating a subject having cancerusing one or more of the methods provided herein, together withradiation therapy. Particular embodiments provide treating a subjecthaving cancer using one or more of the methods provided herein, togetherwith two or more of the treatments selected from surgery, chemotherapy,immunotherapy, targeted therapy, and radiation therapy. Particularembodiments provide treating a subject having cancer using one or moreof the methods provided herein, together with two or more of thetreatments selected from surgery, chemotherapy, and radiation therapy.

In certain embodiments, the subject to be treated with one of themethods provided herein has not been treated with anticancer therapyprior to the administration of the cytidine analog. In certainembodiments, the subject to be treated with one of the methods providedherein has been treated with one or more anticancer therapies prior tothe administration of the cytidine analog. In certain embodiments, thesubject to be treated with one of the methods provided herein has beentreated with a cancer therapeutic agent, as described herein. In certainembodiments, the subject to be treated with one of the methods providedherein has developed drug resistance to anticancer therapy. In certainembodiments, the subject to be treated with the methods provided hereinhas a relapsed cancer. In certain embodiments, the subject to be treatedwith the methods provided herein has a refractory cancer. In certainembodiments, the subject to be treated with the methods provided hereinhas a metastatic cancer.

In one embodiment, the methods provided herein encompass treating asubject regardless of patient's age, although some diseases or disordersare more common in certain age groups. Further provided herein is amethod for treating a subject who has undergone surgery in an attempt totreat the disease or condition at issue. Further provided herein is amethod for treating a subject who has not undergone surgery as anattempt to treat the disease or condition at issue. Because the subjectswith cancer have heterogeneous clinical manifestations and varyingclinical outcomes, the treatment given to a particular subject may vary,depending on his/her prognosis. The skilled clinician will be able toreadily determine without undue experimentation, specific secondaryagents, types of surgery, and types of non-drug based standard therapythat can be effectively used to treat an individual subject with cancer.

In each embodiment provided herein, the method may further comprise oneor more diagnostic steps, to determine, e.g., the type of cancer, thepresence of particular cell types, the genetic profile of a subject,and/or the staging of the disease in a subject.

In each embodiment provided herein, the method may further comprise adisease evaluation step after the cytidine analog has been administeredto the subject, to determine, e.g., changes in one or more molecularmarkers as described herein elsewhere, changes in tumor size andlocation, and/or other benchmarks used by those skilled in the art todetermine the prognosis of cancer in a subject.

Certain methods herein provide administration of a cytidine analog by,e.g., intravenous (IV), subcutaneous (SC) or oral routes ofadministration. Certain methods herein provide administration of acytidine analog by oral route of administration. Certain embodimentsherein provide co-administration of a cytidine analog (e.g.,5-azacytidine or another cytidine analog provided herein) with one ormore additional active agents to provide a synergistic therapeuticeffect in subjects in need thereof. The co-administered agent(s) may bea cancer therapeutic agent, as described herein. In certain embodiments,the co-administered agent(s) may be dosed, e.g., orally or by injection(e.g., IV or SC).

Certain embodiments herein provide methods for treating disorders ofabnormal cell proliferation comprising administering a cytidine analogusing, e.g., IV, SC and/or oral administration methods. Certainembodiments herein provide methods for treating disorders of abnormalcell proliferation comprising administering a cytidine analog using oraladministration methods. In certain embodiments, treatment cyclescomprise multiple doses administered to a subject in need thereof overmultiple days (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, or greater than 28days), optionally followed by treatment dosing holidays (e.g., 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,23, 24, 25, 26, 27, 28, or greater than 28 days). Suitable dosageamounts for the methods provided herein include, e.g., therapeuticallyeffective amounts and prophylactically effective amounts. In specificembodiments, a treatment cycle comprises multiple doses administered toa subject in need thereof once a day or more than once a day, for 3days, for 5 days, for 7 days, for 14 days, for 21 days, or for 28 days.In specific embodiments, a treatment cycle comprises a resting period of1 day, 2 days, 3 days, 4 days, 5 days, 7 days, 14 days, 21 days, or 28days. In specific embodiments, a subject is treated with multipletreatment cycles, for example, multiple 7-day, 14-day, 21-day, 28-day,35-day, or 42-day treatment cycles for a total period of treatment ofabout 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 21, 22, 23, or 24 months, or greater than 24 months. In specificembodiments, a subject is treated with multiple treatment cycles, thatmay be the same or different (e.g., a 7-day treatment cycle followed bya 14-day, 21-day, or 28-day treatment cycle).

In one embodiment, the amount of the cytidine analog (e.g.,5-azacytidine or another cytidine analog provided herein) administeredin the methods provided herein may range, e.g., between about 50mg/m²/day and about 2,000 mg/m²/day, between about 100 mg/m²/day andabout 1,000 mg/m²/day, between about 50 mg/m²/day and about 200mg/m²/day, between about 50 mg/m²/day and about 100 mg/m²/day, betweenabout 100 mg/m²/day and about 500 mg/m²/day, or between about 120mg/m²/day and about 250 mg/m²/day. In certain embodiments, particulardosages are, e.g., about 50 mg/m²/day, about 75 mg/m²/day, about 100mg/m²/day, about 120 mg/m²/day, about 140 mg/m²/day, about 150mg/m²/day, about 180 mg/m²/day, about 200 mg/m²/day, about 220mg/m²/day, about 240 mg/m²/day, about 250 mg/m²/day, about 260mg/m²/day, about 280 mg/m²/day, about 300 mg/m²/day, about 320mg/m²/day, about 350 mg/m²/day, about 380 mg/m²/day, about 400mg/m²/day, about 450 mg/m²/day, or about 500 mg/m²/day. In certainembodiments, particular dosages are, e.g., up to about 100 mg/m²/day, upto about 120 mg/m²/day, up to about 140 mg/m²/day, up to about 150mg/m²/day, up to about 180 mg/m²/day, up to about 200 mg/m²/day, up toabout 220 mg/m²/day, up to about 240 mg/m²/day, up to about 250mg/m²/day, up to about 260 mg/m²/day, up to about 280 mg/m²/day, up toabout 300 mg/m²/day, up to about 320 mg/m²/day, up to about 350mg/m²/day, up to about 380 mg/m²/day, up to about 400 mg/m²/day, up toabout 450 mg/m²/day, up to about 500 mg/m²/day, up to about 750mg/m²/day, or up to about 1000 mg/m²/day.

In one embodiment, the amount of the cytidine analog (e.g.,5-azacytidine or another cytidine analog provided herein) administeredin the methods provided herein may range, e.g., between about 5 mg/dayand about 2,000 mg/day, between about 10 mg/day and about 2,000 mg/day,between about 20 mg/day and about 2,000 mg/day, between about 50 mg/dayand about 1,000 mg/day, between about 100 mg/day and about 600 mg/day,between about 100 mg/day and about 500 mg/day, between about 150 mg/dayand about 500 mg/day, between about 250 mg/day and about 350 mg/day, orbetween about 150 mg/day and about 250 mg/day. In certain embodiments,particular dosages are, e.g., about 10 mg/day, about 20 mg/day, about 50mg/day, about 75 mg/day, about 100 mg/day, about 120 mg/day, about 150mg/day, about 180 mg/day, about 200 mg/day, about 240 mg/day, about 250mg/day, about 280 mg/day, about 300 mg/day, about 320 mg/day, about 350mg/day, about 360 mg/day, about 400 mg/day, about 450 mg/day, about 500mg/day, about 600 mg/day, about 700 mg/day, about 800 mg/day, about 900mg/day, about 1,000 mg/day, about 1,200 mg/day, or about 1,500 mg/day.In certain embodiments, particular dosages are, e.g., up to about 10mg/day, up to about 20 mg/day, up to about 50 mg/day, up to about 75mg/day, up to about 100 mg/day, up to about 120 mg/day, up to about 150mg/day, up to about 200 mg/day, up to about 250 mg/day, up to about 300mg/day, up to about 350 mg/day, up to about 400 mg/day, up to about 450mg/day, up to about 500 mg/day, up to about 600 mg/day, up to about 700mg/day, up to about 800 mg/day, up to about 900 mg/day, up to about1,000 mg/day, up to about 1,200 mg/day, or up to about 1,500 mg/day.

In one embodiment, the amount of the cytidine analog (e.g.,5-azacytidine or another cytidine analog provided herein) in thepharmaceutical composition or dosage form provided herein may range,e.g., between about 5 mg and about 2,000 mg, between about 10 mg andabout 2,000 mg, between about 20 mg and about 2,000 mg, between about 50mg and about 1,000 mg, between about 100 mg and about 600 mg, betweenabout 100 mg and about 500 mg, between about 150 mg and about 500 mg,between about 250 mg and about 350 mg, or between about 150 mg and about250 mg. In certain embodiments, particular amounts are, e.g., about 10mg, about 20 mg, about 50 mg, about 75 mg, about 100 mg, about 120 mg,about 150 mg, about 180 mg, about 200 mg, about 240 mg, about 250 mg,about 300 mg, about 320 mg, about 350 mg, about 360 mg, about 400 mg,about 420 mg, about 450 mg, about 480 mg, about 500 mg, about 600 mg,about 700 mg, about 800 mg, about 900 mg, about 1,000 mg, about 1,200mg, or about 1,500 mg. In certain embodiments, particular amounts are,e.g., up to about 10 mg, up to about 20 mg, up to about 50 mg, up toabout 75 mg, up to about 100 mg, up to about 120 mg, up to about 150 mg,up to about 200 mg, up to about 250 mg, up to about 300 mg, up to about350 mg, up to about 400 mg, up to about 450 mg, up to about 500 mg, upto about 600 mg, up to about 700 mg, up to about 800 mg, up to about 900mg, up to about 1,000 mg, up to about 1,200 mg, or up to about 1,500 mg.

In one embodiment, depending on the disease to be treated and thesubject's condition, the cytidine analog (e.g., 5-azacytidine or anothercytidine analog provided herein) may be administered by oral, parenteral(e.g., intramuscular, intraperitoneal, intravenous, CIV, intracistemalinjection or infusion, subcutaneous injection, or implant), inhalation,nasal, vaginal, rectal, sublingual, or topical (e.g., transdermal orlocal) routes of administration. In some embodiments, the cytidineanalog may be formulated, alone or together with one or more activeagent(s), in suitable dosage unit with pharmaceutically acceptableexcipients, carriers, adjuvants and vehicles, appropriate for each routeof administration. In one embodiment, the cytidine analog (e.g.,5-azacytidine or another cytidine analog provided herein) isadministered orally. In another embodiment, the cytidine analog (e.g.,5-azacytidine or another cytidine analog provided herein) isadministered parenterally. In yet another embodiment, the cytidineanalog (e.g., 5-azacytidine or another cytidine analog provided herein)is administered intravenously. In yet another embodiment, the cytidineanalog (e.g., 5-azacytidine or another cytidine analog provided herein)is administered subcutaneously.

In one embodiment, the cytidine analog (e.g., 5-azacytidine or anothercytidine analog provided herein) can be delivered as a single dose suchas, e.g., a single bolus injection, or oral tablets or pills; or overtime such as, e.g., continuous infusion over time or divided bolus dosesover time. In one embodiment, the cytidine analog (e.g., 5-azacytidineor another cytidine analog provided herein) can be administeredrepetitively if necessary, for example, until the patient experiencesstable disease or regression, or until the patient experiences diseaseprogression or unacceptable toxicity. For example, stable disease forsolid tumors generally means that the perpendicular diameter ofmeasurable lesions has not increased by 25% or more from the lastmeasurement. See, e.g., Response Evaluation Criteria in Solid Tumors(RECIST) Guidelines, Journal of the National Cancer Institute 92(3):205-216 (2000). Stable disease or lack thereof is determined by methodsknown in the art such as evaluation of patient's symptoms, physicalexamination, visualization of the tumor that has been imaged usingX-ray, CAT, PET, or MRI scan and other commonly accepted evaluationmodalities.

In one embodiment, the cytidine analog (e.g., 5-azacytidine or anothercytidine analog provided herein) can be administered once daily (QD), ordivided into multiple daily doses such as twice daily (BID), three timesdaily (TID), and four times daily (QID). In one embodiment, theadministration can be continuous (i.e., daily for consecutive days orevery day), intermittent, e.g., in cycles (i.e., including days, weeks,or months of rest when no drug is administered). In one embodiment, thecytidine analog is administered daily, for example, once or more thanonce each day for a period of time. In one embodiment, the cytidineanalog is administered daily for an uninterrupted period of at least 7days, in some embodiments, up to 52 weeks. In one embodiment, thecytidine analog is administered intermittently, i.e., stopping andstarting at either regular or irregular intervals. In one embodiment,the cytidine analog is administered for one to six days per week. In oneembodiment, the cytidine analog is administered in cycles (e.g., dailyadministration for about one, two, three, four, five, six, seven, oreight consecutive weeks, then a rest period with no administration forabout one, two, three, or four weeks). In one embodiment, the cytidineanalog is administered on alternate days. In one embodiment, thecytidine analog is administered in cycles (e.g., administered daily orcontinuously for a certain period interrupted with a rest period).

In one embodiment, the frequency of administration ranges from aboutdaily to about monthly. In certain embodiments, the cytidine analog(e.g., 5-azacytidine or another cytidine analog provided herein) isadministered once a day, twice a day, three times a day, four times aday, once every other day, twice a week, once every week, once every twoweeks, once every three weeks, or once every four weeks. In oneembodiment, the cytidine analog is administered once a day. In anotherembodiment, the cytidine analog is administered twice a day. In yetanother embodiment, the cytidine analog is administered three times aday. In still another embodiment, the cytidine analog is administeredfour times a day.

In one embodiment, the cytidine analog (e.g., 5-azacytidine or anothercytidine analog provided herein) is administered once per day from oneday to six months, from one week to three months, from one week to fourweeks, from one week to three weeks, or from one week to two weeks. Incertain embodiments, the cytidine analog is administered once per dayfor one week, two weeks, three weeks, or four weeks. In one embodiment,the cytidine analog is administered once per day for one week. Inanother embodiment, the cytidine analog is administered once per day fortwo weeks. In yet another embodiment, the cytidine analog isadministered once per day for three weeks. In still another embodiment,the cytidine analog is administered once per day for four weeks.

In one embodiment, the cytidine analog (e.g., 5-azacytidine or anothercytidine analog provided herein) is administered once per day for about1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 6 weeks,about 9 weeks, about 12 weeks, about 15 weeks, about 18 weeks, about 21weeks, or about 26 weeks. In certain embodiments, the cytidine analog isadministered intermittently. In certain embodiments, the cytidine analogis administered intermittently in the amount of between about 50mg/m²/day and about 2,000 mg/m²/day. In certain embodiments, thecytidine analog is administered intermittently in the amount of betweenabout 100 mg/day and about 600 mg/day. In certain embodiments, thecytidine analog is administered continuously. In certain embodiments,the cytidine analog is administered continuously in the amount ofbetween about 50 mg/m²/day and about 1,000 mg/m²/day. In certainembodiments, the cytidine analog is administered continuously in theamount of between about 100 mg/day and about 600 mg/day.

In certain embodiments, the cytidine analog (e.g., 5-azacytidine oranother cytidine analog provided herein) is administered to a patient incycles. Cycling therapy involves the administration of an active agentfor a period of time, followed by a rest for a period of time, andrepeating this sequential administration. Cycling therapy can reduce thedevelopment of resistance, avoid or reduce the side effects, and/orimproves the efficacy of the treatment.

In one embodiment, the cytidine analog (e.g., 5-azacytidine or anothercytidine analog provided herein) is administered daily in single ordivided doses for about 3 days, about 5 days, about one week, about twoweeks, about three weeks, about four weeks, about five weeks, about sixweeks, about seven weeks, about eight weeks, about ten weeks, aboutfifteen weeks, or about twenty weeks, followed by a rest period of about1 day to about ten weeks. In one embodiment, the methods provided hereincontemplate cycling treatments of about one week, about two weeks, aboutthree weeks, about four weeks, about five weeks, about six weeks, abouteight weeks, about ten weeks, about fifteen weeks, or about twentyweeks. In some embodiments, the cytidine analog is administered daily insingle or divided doses for about 3 days, about 5 days, about one week,about two weeks, about three weeks, about four weeks, about five weeks,or about six weeks with a rest period of about 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 29, or 30 days. In someembodiments, the rest period is 1 day. In some embodiments, the restperiod is 3 days. In some embodiments, the rest period is 7 days. Insome embodiments, the rest period is 14 days. In some embodiments, therest period is 28 days. The frequency, number and length of dosingcycles can be increased or decreased.

In one embodiment, the methods provided herein comprise: i)administering to the subject a first daily dose of a cytidine analog;ii) optionally resting for a period of at least one day where thecytidine analog is not administered to the subject; iii) administering asecond dose of the cytidine analog to the subject; and iv) repeatingsteps ii) to iii) a plurality of times. In certain embodiments, thefirst daily dose is between about 50 mg/m²/day and about 2,000mg/m²/day. In certain embodiments, the second daily dose is betweenabout 50 mg/m²/day and about 2,000 mg/m²/day. In certain embodiments,the first daily dose is between about 100 mg/day and about 1,000 mg/day.In certain embodiments, the second daily dose is between about 100mg/day and about 1,000 mg/day. In certain embodiments, the first dailydose is higher than the second daily dose. In certain embodiments, thesecond daily dose is higher than the first daily dose. In oneembodiment, the rest period is 1 day, 2 days, 3 days, 5 days, 7 days, 10days, 12 days, 13 days, 14 days, 15 days, 17 days, 21 days, or 28 days.

In certain embodiments, the cytidine analog (e.g., 5-azacytidine oranother cytidine analog provided herein) is administered continuouslyfor between about 1 and about 52 weeks. In certain embodiments, thecytidine analog is administered continuously for about 0.5, 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, or 12 months. In certain embodiments, thecytidine analog is administered continuously for about 7, about 14,about 21, about 28, about 35, about 42, about 84, or about 112 days. Itis understood that the duration of the treatment may vary with the age,weight, and condition of the subject being treated, and may bedetermined empirically using known testing protocols or according to theprofessional judgment of the person providing or supervising thetreatment. The skilled clinician will be able to readily determine,without undue experimentation, an effective drug dose and treatmentduration, for treating an individual subject having a particular type ofcancer.

1. Methods of Using Oral Formulations Provided Herein

As described herein, certain embodiments herein provide oralformulations of cytidine analogs useful in methods relating to, e.g.,permitting different dosing amounts and/or dosing periods; providingalternative pharmacokinetic profiles, pharmacodynamic profiles, and/orsafety profiles; permitting the evaluation of long-term and/ormaintenance therapies; providing treatment regimens that maximizedemethylation and/or gene re-expression; providing treatment regimensthat prolong continuous demethylation; providing new indications forcytidine analogs; and/or providing other potential advantageousbenefits.

Provided herein are methods of treating, preventing, or managingpatho-physiological conditions manifested by abnormal cellproliferation, such as, for example, cancer, including hematologicaldisorders and solid tumors, by orally administering a pharmaceuticalformulation comprising a cytidine analog, such as, for example,5-azacytidine, wherein the formulation releases the cytidine analogsubstantially in the stomach. Other embodiments herein provide methodsof treating, preventing, or managing immune disorders. In particularembodiments, the methods provided herein involve oral administering aformulation that effects an immediate release of the cytidine analog. Incertain embodiments, the cytidine analog and one or more therapeuticagents are co-administered to subjects to yield a synergistictherapeutic effect. The co-administered agent may be a cancertherapeutic agent dosed orally or by injection.

In certain embodiments, methods provided herein for treating,preventing, or managing disorders related to abnormal cell proliferationcomprise orally administering a formulation comprising a therapeuticallyeffective amount of a cytidine analog. Particular therapeuticindications relating to the methods provided herein are disclosedherein. In certain embodiments, the therapeutically effective amount ofthe cytidine analog in the pharmaceutical formulation is an amount asdisclosed herein. In certain embodiments, the precise therapeuticallyeffective amount of the cytidine analog in the pharmaceuticalformulation will vary depending on, e.g., the age, weight, diseaseand/or condition of the subject.

In particular embodiments, the disorders related to abnormal cellproliferation include, but are not limited to, solid tumors, sarcoma,melanoma, carcinoma, adenocarcinoma, chordoma, breast cancer, colorectalcancer, ovarian cancer, lung cancer (e.g., non-small-cell lung cancerand small-cell lung cancer), testicular cancer, renal cancer, bladdercancer, pancreatic cancer, bone cancer, gastric cancer, head and neckcancer, prostate cancer, MDS, AML, ALL, CML, leukemia, chroniclymphocytic leukemia (CLL), lymphoma (including non-Hodgkin's lymphoma(NHL) and Hodgkin's lymphoma), and multiple myeloma (MM). In particularembodiments, the disorder related to abnormal cell proliferation is asolid tumor. In particular embodiments, the disorder related to abnormalcell proliferation is a relapsed or refractory solid tumor. Inparticular embodiments, the disorder related to abnormal cellproliferation is MDS. In particular embodiments, the disorder related toabnormal cell proliferation is AML. In particular embodiments, thedisorder related to abnormal cell proliferation is breast cancer. Inparticular embodiments, the disorder related to abnormal cellproliferation is bladder cancer. In particular embodiments, the disorderrelated to abnormal cell proliferation is head and neck cancer. Inparticular embodiments, the disorder related to abnormal cellproliferation is pancreatic cancer. In particular embodiments, thedisorder related to abnormal cell proliferation is lung cancer (e.g.,NSCLC or SCLC). In particular embodiments, the disorder related toabnormal cell proliferation is ovarian cancer. In particularembodiments, the disorder related to abnormal cell proliferation iscolorectal cancer. In particular embodiments, the disorder related toabnormal cell proliferation is skin cancer (e.g., melanoma). Inparticular embodiments, the disorder related to abnormal cellproliferation is uterine cancer. In particular embodiments, the disorderrelated to abnormal cell proliferation is sarcoma.

In one embodiment, methods provided herein for treating, preventing, ormanaging disorders of abnormal cell proliferation comprise administeringa cytidine analog orally. In other embodiments, methods provided hereinfor treating, preventing, or managing disorders of abnormal cellproliferation comprise administering a cytidine analog using at leasttwo of IV, SC and oral administration methods. For example, particularembodiments herein provide administering an initial treatment cycle of acytidine analog, such as, for example, 5-azacytidine, administeredeither SC or IV, followed by subsequent orally administered treatmentcycles of the cytidine analog. In certain embodiments, treatment cyclescomprise multiple doses administered to a subject in need thereof overmultiple days (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19, 20, 21, or greater than 21 days), optionally followed bytreatment dosing holidays (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, or greater than 14 days). Particular embodiments herein providea treatment schedule comprising SC and/or IV administration for one,two, three, four, five, or more initial cycles, followed by oraladministration for subsequent cycles. For example, particularembodiments herein provide a treatment schedule comprising SCadministration for cycle 1, followed by oral administration forsubsequent cycles. Suitable dosage ranges and amounts for the methodsprovided herein are provided throughout the specification. For example,in certain embodiments, the SC dose is about 75 mg/m². In certainembodiments, the oral dose is about 50 mg, about 60 mg, about 70 mg,about 80 mg, about 90 mg, about 100 mg, about 120 mg, about 140 mg,about 150 mg, about 160 mg, about 180 mg, about 200 mg, about 220 mg,about 240 mg, about 250 mg, about 260 mg, about 280 mg, about 300 mg,about 320 mg, about 340 mg, about 350 mg, about 360 mg, about 380 mg,about 400 mg, about 420 mg, about 450 mg, about 480 mg, about 500 mg,about 600 mg, or greater than about 600 mg. In certain embodiments, oraldoses are calculated to achieve about 80%, 100%, or 120% of SC AUC.

In certain embodiments, methods of treating disorders of abnormal cellproliferation comprises orally administering a formulation comprising acytidine analog (e.g., 5-azacytidine or another cytidine analog providedherein) as single or multiple daily doses. In particular embodiments, aformulation comprising a cytidine analog is orally administered once perday, twice per day, three times per day, four times per day, or morethan four times per day. For example, in certain embodiments, aformulation comprising a cytidine analog is administered using atreatment cycle comprising administration of about 50 mg, about 60 mg,about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 120 mg, about140 mg, about 150 mg, about 160 mg, about 180 mg, about 200 mg, about220 mg, about 240 mg, about 250 mg, about 260 mg, about 280 mg, about300 mg, about 320 mg, about 340 mg, about 350 mg, about 360 mg, about380 mg, about 400 mg, about 420 mg, about 450 mg, about 480 mg, about500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, or about1,000 mg of the cytidine analog once, twice, three, or four times perday for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 days. In certainembodiments, the method of treating comprises continuous low-doseadministration. In certain embodiments, the formulation comprising thecytidine analog is administered using a treatment cycle comprisingadministration of about 200 mg of the cytidine analog once per day for 7or more days. In certain embodiments, the formulation comprising thecytidine analog is administered using a treatment cycle comprisingadministration of about 200 mg of the cytidine analog twice per day for7 or more days. In certain embodiments, the formulation comprising thecytidine analog is administered using a treatment cycle comprisingadministration of about 200 mg of the cytidine analog once per day for14 or more days. In certain embodiments, the formulation comprising thecytidine analog is administered using a treatment cycle comprisingadministration of about 200 mg of the cytidine analog twice per day for14 or more days. In certain embodiments, the formulation comprising thecytidine analog is administered using a treatment cycle comprisingadministration of about 200 mg of the cytidine analog once per day for21 or more days. In certain embodiments, the formulation comprising thecytidine analog is administered using a treatment cycle comprisingadministration of about 200 mg of the cytidine analog twice per day for21 or more days. In certain embodiments, the formulation comprising thecytidine analog is administered using a treatment cycle comprisingadministration of about 200 mg of the cytidine analog three times perday for 7 or more days. In certain embodiments, the formulationcomprising the cytidine analog is administered using a treatment cyclecomprising administration of about 200 mg of the cytidine analog threetimes per day for 14 or more days. In certain embodiments, theformulation comprising the cytidine analog is administered using atreatment cycle comprising administration of about 300 mg of thecytidine analog once per day for 7 or more days. In certain embodiments,the formulation comprising the cytidine analog is administered using atreatment cycle comprising administration of about 300 mg of thecytidine analog twice per day for 7 or more days. In certainembodiments, the formulation comprising the cytidine analog isadministered using a treatment cycle comprising administration of about300 mg of the cytidine analog once per day for 14 or more days. Incertain embodiments, the formulation comprising the cytidine analog isadministered using a treatment cycle comprising administration of about300 mg of the cytidine analog twice per day for 14 or more days. Incertain embodiments, the formulation comprising the cytidine analog isadministered using a treatment cycle comprising administration of about300 mg of the cytidine analog once per day for 21 or more days. Incertain embodiments, the formulation comprising the cytidine analog isadministered using a treatment cycle comprising administration of about300 mg of the cytidine analog twice per day for 21 or more days. Incertain embodiments, the formulation comprising the cytidine analog isadministered using a treatment cycle comprising administration of about300 mg of the cytidine analog three times per day for 7 or more days. Incertain embodiments, the formulation comprising the cytidine analog isadministered using a treatment cycle comprising administration of about300 mg of the cytidine analog three times per day for 14 or more days.In certain embodiments, methods provided herein comprise administering aformulation comprising a cytidine analog using one or more of the cyclesprovided herein, and repeating one or more of the cycles for a periodof, e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or greater than12 months.

In certain embodiments, methods herein comprise administering particularoral formulations provided herein to, e.g., overcome limitationsassociated with IV or SC administration of cytidine analogs. Forexample, IV or SC administration may limit the ability to deliver acytidine analog for longer periods of time on a regular basis, therebypotentially limiting the maximal efficacy of the cytidine analog. Due tothe difficulties of complying with the rigors of a prolonged IV or SCdosing schedule, prolonged SC or IV exposure to a cytidine analog maycause subjects (e.g., subjects with multiple cytopenias) to discontinuefrom the regimen. See, e.g., Lyons, R. M., et al., Hematologic Responseto Three Alternative Dosing Schedules of Azacitidine in Patients WithMyelodysplastic Syndromes, J. Clin. Oncol. (2009)(DOI:10.1200/JCO.2008.17.1058), which is incorporated by referenceherein in its entirety. Accordingly, in certain embodiments, methodsprovided herein comprise administering an oral formulation providedherein to overcome these or other limitations associated with SC or IVcytidine analog administration. For example, in certain embodiments,methods provided herein comprise administering daily to a subject anoral formulation provided herein for 7 or more, 8 or more, 9 or more, 10or more, 11 or more, 12 or more, 13 or more, 14 or more, 15 or more, 16or more, 17 or more, 18 or more, 19 or more, 20 or more, 21 or more, or28 or more days.

Certain embodiments herein provide methods comprising administering oralformulations of cytidine analogs provided herein comprising deliveringthe cytidine analog (e.g., 5-azacytidine or another cytidine analogprovided herein) at a lower dose over a more prolonged period of time,as compared to IV or SC administration. In particular embodiments, suchmethods comprise managing dose-related cytopenias (including, e.g.,dose-related cytopenias associated with 5-azacytidine) by administeringan oral formulation provided herein. In certain embodiments, methodsprovided herein comprise administering an oral formulation providedherein to achieve an improved safety profile as compared to an IV or SCdose comprising the same cytidine analog.

As described herein, certain embodiments provide methods for improvedtreatment of particular diseases or disorders (e.g., treatment of solidtumors) by administering an oral formulation provided herein, ascompared to IV or SC administration of the cytidine analog. Inparticular embodiments, certain methods herein provide administeringoral formulations provided herein at lower doses for more prolongedperiods of time, leading to improved demethylation. For example, certainmethods provided herein comprise administering an oral formulationprovided herein to treat a solid tumor while avoiding certaindose-limiting-toxicity-related side effects associated with dosing thecytidine analog via SC or IV administration. An example of certaintoxicity-related drawbacks associated with administration of a cytidineanalog are described, e.g., in K. Appleton et al., J. Clin. Oncol., Vol.25(29):4603-4609 (2007), which is incorporated by reference herein inits entirety.

Particular embodiments herein provide methods for treating a subjecthaving a disease or disorder provided herein by orally administering apharmaceutical composition provided herein, wherein the treatmentresults in improved survival of the subject. In certain embodiments, theimproved survival is measured as compared to one or more conventionalcare regimens. Particular embodiments herein provide methods fortreating a subject having a disease or disorder provided herein byorally administering a pharmaceutical composition provided herein,wherein the treatment provides improved effectiveness. In particularembodiments, the improved effectiveness is measured using one or moreendpoints for cancer clinical trials, as recommended by the U.S. Foodand Drug Administration (FDA). For example, FDA provides Guidance forIndustry on Clinical Trial Endpoints for the Approval of Cancer Drugsand Biologics (http://www.fda.gov/CbER/gdlns/clintrialend.htm). The FDAendpoints include, but are not limited to, Overall Survival, EndpointsBased on Tumor Assessments such as (i) Disease-Free Survival (ii)Objective Response Rate, (iii) Time to Progression and Progression-FreeSurvival and (iv) Time-to-Treatment Failure. Endpoints Involving SymptomEndpoints may include Specific Symptom Endpoints such as (i) Time toprogression of cancer symptoms and (ii) A composite symptom endpoint.Biomarkers assayed from blood or body fluids may also be useful todetermine the management of the disease.

In certain embodiments, the methods of treating disorders of abnormalcell proliferation comprise orally administering a formulation of acytidine analog with food. In certain embodiments, the methods oftreating disorders of abnormal cell proliferation comprise orallyadministering a formulation of a cytidine analog without food. Incertain embodiments, pharmacological parameters (e.g., Cmax, Tmax)depend on the fed state of the subject. In certain embodiments, theformulation of the cytidine analog is administered sublingually.

In certain embodiments, the cytidine analog, e.g., 5-azacytidine oranother cytidine analog provided herein, is not co-administered with acytidine deaminase inhibitor. In certain embodiments, the oralformulation comprising a cytidine analog as provided herein is notco-administered with THU. Certain embodiments herein provide methods oftreating a disease or disorder provided herein (e.g., a diseaseassociated with abnormal cell proliferation) comprising orallyadministering a cytidine analog provided herein for releasesubstantially in the stomach, wherein the methods achieve a particularbiological parameter provided herein (e.g., a particular Cmax value,Tmax value, and/or AUC value provided herein), and wherein the methodscomprise not co-administering a cytidine deaminase inhibitor with thecytidine analog. Certain embodiments herein provide methods of treatinga disease or disorder provided herein (e.g., a disease associated withabnormal cell proliferation) comprising orally administering a cytidineanalog provided herein for release substantially in the stomach, whereinthe methods avoid adverse effects associated with administering acytidine deaminase inhibitor (e.g., THU) by not co-administering thecytidine deaminase inhibitor with the cytidine analog. In particularembodiments, a cytidine deaminase inhibitor (e.g., THU) isco-administered with the cytidine analog in an amount of, e.g., lessthan about 500 mg/d, less than about 200 mg/d, less than about 150 mg/d,less than about 100 mg/d, less than about 50 mg/d, less than about 25mg/d, less than about 10 mg/d, less than about 5 mg/d, less than about 1mg/d, or less than about 0.1 mg/d.

Certain embodiments herein provide methods for delivering a cytidineanalog to a subject comprising administering to the subject in needthereof an oral formulation comprising a cytidine analog. In particularembodiments, oral formulations comprise (1) a therapeutically effectiveamount of a cytidine analog; and (2) an optional drug releasecontrolling component capable of releasing the cytidine analogsubstantially in the stomach after a subject ingests the oralformulation comprising the cytidine analog. Certain embodiments hereinprovide a method for enhancing the oral bioavailability of a cytidineanalog in a subject. Certain embodiments herein provide a method ofincreasing the oral bioavailability of a cytidine analog comprisingorally administering a pharmaceutical composition provided herein. Incertain methods provided herein, a pharmaceutical composition providedherein is orally administered to a subject, contacts the biologicalfluids of the subject's body, and is absorbed in the uppergastrointestinal tract, such as, for example, substantially in thestomach.

Certain embodiments herein provide a method of achieving a particularexposure value provided herein by administering an oral formulationcomprising a cytidine analog provided herein. Certain embodiments hereinprovide a method of achieving a particular oral bioavailability valueprovided herein by administering an oral formulation comprising acytidine analog provided herein. Certain embodiments herein provide amethod of achieving a particular AUC value provided herein byadministering an oral formulation comprising a cytidine analog providedherein. Certain embodiments herein provide a method of achieving aparticular Cmax value provided herein by administering an oralformulation comprising a cytidine analog provided herein. Certainembodiments herein provide a method of achieving a particular Tmax valueprovided herein by administering an oral formulation comprising acytidine analog provided herein.

Certain embodiments herein provide methods of treating a conditioninvolving undesirable or uncontrolled cell proliferation byadministering an oral formulation comprising a cytidine analog asprovided herein. Such conditions include, e.g., benign tumors, varioustypes of cancers such as primary tumors and tumor metastasis, solidtumors (e.g., relapsed or refractory solid tumors), hematologicaldisorders (e.g. leukemia, myelodysplastic syndrome and sickle cellanemia), restenosis (e.g. coronary, carotid, and cerebral lesions),abnormal stimulation of endothelial cells (arteriosclerosis), insults tobody tissue due to surgery, abnormal wound healing, abnormalangiogenesis, diseases that produce fibrosis of tissue, repetitivemotion disorders, disorders of tissues that are not highly vascularized,and proliferative responses associated with organ transplants.

In certain embodiments, cells in a benign tumor retain theirdifferentiated features and do not divide in a completely uncontrolledmanner. A benign tumor may be localized and/or nonmetastatic. Specifictypes of benign tumors that can be treated using the methods,compositions, and formulations provided herein include, e.g.,hemangiomas, hepatocellular adenoma, cavernous hemangioma, focal nodularhyperplasia, acoustic neuromas, neurofibroma, bile duct adenoma, bileduct cystanoma, fibroma, lipomas, leiomyomas, mesotheliomas, teratomas,myxomas, nodular regenerative hyperplasia, trachomas and pyogenicgranulomas.

In certain embodiments, cells in a malignant tumor becomeundifferentiated, do not respond to the body's growth control signals,and/or multiply in an uncontrolled manner. The malignant tumor may beinvasive and capable of spreading to distant sites (metastasizing).Malignant tumors may be divided into two categories: primary andsecondary. Primary tumors arise directly from the tissue in which theyare found. A secondary tumor, or metastasis, is a tumor which isoriginated elsewhere in the body but has now spread to a distant organ.The common routes for metastasis are direct growth into adjacentstructures, spread through the vascular or lymphatic systems, andtracking along tissue planes and body spaces (peritoneal fluid,cerebrospinal fluid, etc.).

Without being limited by a particular theory, methylation can lead tothe silencing of genes critical to cellular control (i.e., epigeneticgene silencing), and can be an early event in the development ofmalignant tumors including, e.g., colorectal cancer or lung cancer. See,e.g., M. V. Brock et al., N. Engl. J. Med., 2008, 358(11):1118-28; P. M.Das et al., Mol. Cancer, 2006, 5(28); G. Gifford et al., Clin. CancerRes., 2004, 10:4420-26; J. G. Herman et al., N. Engl. J. Med., 2003,349:2042-54; A. M. Jubb et al., J. Pathology, 2001, 195:111-34.Accordingly, in certain embodiments, without being limited by aparticular theory, methods herein provide using oral formulationsprovided herein to prevent or reverse epigenetic gene silencing, e.g.,by reversing abnormal DNA methylation. In specific embodiments, oralformulations provided herein are used for early intervention to preventthe development of cancer in patients at risk of developing cancer,e.g., familial polyposis or lung cancer, wherein a cause of the canceris epigenetic gene silencing. In particular embodiments, such earlyintervention would be impractical by means other than oraladministration (e.g., IV or SC administration). In specific embodiments,oral formulations provided herein are used for early intervention toprevent the recurrence of cancer in patients at risk for early relapse,e.g., colorectal cancer or non-small-cell lung cancer. In certainembodiments, the early intervention is achieved via prolonged oraldosing schedules, using formulations and/or methods as described herein.Certain embodiments provide methods for administering oral formulationsprovided herein to reverse the effect of gene silencing, e.g., inpatients at risk of gene silencing due to epigenetic changes.

In certain embodiments, specific types of cancers or malignant tumors,either primary or secondary, that can be treated using the methods,compositions, and formulations provided herein include, e.g., leukemia,lymphoma, breast cancer, skin cancer, bone cancer, prostate cancer,liver cancer, lung cancer (e.g., non-small-cell lung cancer andsmall-cell lung cancer), brain cancer, cancer of the larynx, gallbladder, pancreas, rectum, uterine, prostate, parathyroid, thyroid,adrenal, neural tissue, head and neck, colon, stomach, bronchi, kidney,or bladder, basal cell carcinoma, squamous cell carcinoma of bothulcerating and papillary type, melanoma, metastatic skin carcinoma,sarcoma, osteo sarcoma, Ewing's sarcoma, veticulum cell sarcoma,myeloma, multiple myeloma, giant cell tumor, gallstones, islet celltumor, primary brain tumor, acute and chronic lymphocytic andgranulocytic tumors, hairy-cell tumor, adenoma, hyperplasia, medullarycarcinoma, pheochromocytoma, mucosal neuronmas, intestinalganglioneuromas, hyperplastic corneal nerve tumor, marfanoid habitustumor, Wilm's tumor, seminoma, ovarian cancer, leiomyoma tumor, cervicalsquamous cell carcinoma, cervical dysplasia and in situ carcinoma,neuroblastoma, retinoblastoma, medulloblastoma, soft tissue sarcoma,malignant carcinoid, topical skin lesion, mycosis fungoides,rhabdomyosarcoma, Kaposi's sarcoma, osteogenic and other sarcoma,malignant hypercalcemia, renal cell tumor, polycythermia vera,adenocarcinoma, glioblastoma multiforma, malignant melanoma, epidermoidcarcinoma, other carcinomas and sarcomas, relapsed or refractory solidtumors, and advanced metastatic solid tumors.

In certain embodiments, specific types of cancers or malignant tumors,either primary or secondary, that can be treated using the methods,compositions, and formulations provided herein include, e.g., Burkitt'slymphoma, Hodgkin's lymphoma, nasopharyngeal carcinoma, cervicalcarcinoma, head and neck squamous cell carcinoma, Merkel cell tumors,adenocarcinoma of the colon, pancreatic cancer, gall bladderadenocarcinoma, pancreatic adenocarcinoma, sigmoid colon adenocarcinoma,metastatic pancreatic adenocarcinoma, endometrial carcinoma, ovariancancer, pancreatic carcinoma, metastatic ovarian carcinoma,gastrointestinal stromal tumor, metastatic colorectal cancer, breastductal carcinoma, nonsmall cell adenocarcinoma of the lung, papillaryserous endocervical adenocarcinoma, endometroid type well-differentiatedadenocarcinoma, rectal adenocarcinoma, moderately differentiatedadenocarcinoma, metastatic colorectal adenocarcinoma, appendicialmucinous adenocarcinoma, cervical adenocarcinoma, colon Krasadenocarcinoma, and colon adenocarcinoma.

Particular embodiments herein provide using the methods, compositions,and formulations provided herein to treat abnormal cell proliferationdue to, e.g., insults to body tissue during surgery for a variety ofsurgical procedures, including, e.g., joint surgery, bowel surgery, andcheloid scarring. Proliferative responses associated with organtransplantation that may be treated using the methods, compositions, andformulations provided herein include those proliferative responsescontributing to potential organ rejections or associated complications.Specifically, these proliferative responses may occur duringtransplantation of the heart, lung (e.g., non-small-cell lung cancer andsmall-cell lung cancer), liver, kidney, and other body organs or organsystems.

In certain embodiments, the amount of the cytidine analog in the oralformulations provided herein, the methods of administration thereof, orthe methods of treatment as set forth herein, is a specific dosageamount as provided herein.

2. Biomarkers

In certain embodiments, appropriate biomarkers may be used to determineor predict the effect of the methods provided herein on the diseasestate and to provide guidance as to the dosing schedule. For example,particular embodiments herein provide a method for determining whether apatient diagnosed with cancer has an increased probability of obtaininga greater benefit from treatment with a pharmaceutical compositioncomprising a cytidine analog, e.g., by assessing the patient's nucleicacid methylation status. In particular embodiments, the cytidine analogis 5-azacytidine. In particular embodiments, the cytidine analog isdecitabine. In particular embodiments, the nucleic acid is DNA or RNA.In particular embodiments, the greater benefit is an overall survivalbenefit. In particular embodiments, the methylation status is examinedin one or more genes, e.g., genes associated with the particular cancer.Specific embodiments involve methods for determining whether baselineDNA methylation levels influence overall survival in patients withcancer treated with a cytidine analog, such as 5-azacytidine ordecitabine. Specific embodiments provide methods for determining whethergene promoter methylation levels influence overall survival in patientswith cancer.

In one embodiment, provided herein is a method for determining whether apatient diagnosed with cancer has an increased probability of obtaininga greater benefit from treatment with a pharmaceutical compositioncomprising a cytidine analog by assessing the gene expression profile inthe patient. In one embodiment, provided herein is a method fordetermining whether a patient diagnosed with cancer has an increasedprobability of obtaining a greater benefit from treatment with apharmaceutical composition comprising a cytidine analog by assessingmolecular markers, including one or more cell cycle markers, apoptosismarkers, and DNA damage markers. In particular embodiments, the cytidineanalog is 5-azacytidine. In particular embodiments, the cytidine analogis decitabine. In particular embodiments, the greater benefit is anoverall survival benefit.

In certain embodiments, appropriate biomarkers may be used to determineor predict the effect of the pharmaceutical compositions comprisingcytidine analogs on the disease state and to provide guidance to thedosing schedule. For example, particular embodiments herein provide amethod of determining whether a patient diagnosed with a solid tumor,leukemia, lymphoma, multiple myeloma, MDS, or AML, has an increasedprobability of obtaining a greater benefit from treatment with apharmaceutical composition comprising a cytidine analog by assessing thepatient's nucleic acid methylation status. In particular embodiments,the cytidine analog is azacitidine. In particular embodiments, thecytidine analog is decitabine. In particular embodiments, the nucleicacid is DNA or RNA. In particular embodiments, the greater benefit is anoverall survival benefit. In particular embodiments, the methylationstatus is examined in one or more genes, e.g., genes associated with thesolid tumor, leukemia, lymphoma, multiple myeloma, MDS, or AML. Specificembodiments involve methods for determining whether baseline DNAmethylation levels influence overall survival in patients treated withazacitidine. Specific embodiments involve methods for determiningwhether baseline DNA methylation levels influence overall survival inpatients treated with decitabine. Specific embodiments provide methodsfor determining whether gene promoter methylation levels influenceoverall survival in patients.

For example, specific embodiments herein provide methods for evaluatingthe influence of gene methylation on prolonged survival in patients witha solid tumor (e.g., a relapsed or refractory solid tumor). Inparticular embodiments, such evaluation is used to predict overallsurvival in patients with a solid tumor, e.g., upon treatment with apharmaceutical composition comprising a cytidine analog, as providedherein. In particular embodiments, such evaluation is used fortherapeutic decision-making. In specific embodiments, such therapeuticdecision-making includes planning or adjusting a patient's treatment,e.g., the dosing regimen, amount, and/or duration of administration ofthe cytidine analogue.

Certain embodiments provide methods of identifying individual patientsdiagnosed with a solid tumor having an increased probability ofobtaining an overall survival benefit from cytidine analog treatment,using analysis of methylation levels, e.g., in particular genes. In someembodiments, lower levels of nucleic acid methylation are associatedwith an increased probability of obtaining improved overall survivalfollowing treatment with a cytidine analog. In some embodiments, higherlevels of nucleic acid methylation are associated with an increasedprobability of obtaining improved overall survival following treatmentwith a cytidine analog. In some embodiments, a particular pattern orsignature of nucleic acid methylation of multiple genes are associatedwith an increased probability of obtaining improved overall survivalfollowing treatment with a cytidine analog. In some embodiments, theincreased probability of obtaining improved overall survival followingtreatment is at least a 5% greater probability, at least a 10% greaterprobability, at least a 20% greater probability, at least a 30% greaterprobability, at least a 40% greater probability, at least a 50% greaterprobability, at least a 60% greater probability, at least a 70% greaterprobability, at least an 80% greater probability, at least a 90% greaterprobability, at least a 100% greater probability, at least a 125%greater probability, at least a 150% greater probability, at least a175% greater probability, at least a 200% greater probability, at leasta 250% greater probability, at least a 300% greater probability, atleast a 400% greater probability, or at least a 500% greater probabilityof obtaining improved overall survival following treatment, e.g., usinga pharmaceutical composition comprising a cytidine analog as providedherein. In particular embodiments, the greater probability of obtainingimproved overall survival following treatment is a greater probabilityas compared to the average probability of a particular comparisonpopulation of patients.

In particular embodiments, nucleic acid (e.g., DNA or RNA)hypermethylation status may be determined by any method known in theart. In certain embodiments, DNA hypermethylation status may bedetermined using the bone marrow aspirates of patients diagnosed withcancer, e.g., by using quantitative real-time methylation specific PCR(“qMSP”). In certain embodiments, the methylation analysis may involvebisulfite conversion of genomic DNA. For example, in certainembodiments, bisulfite treatment of DNA is used to convertnon-methylated CpG sites to UpG, leaving methylated CpG sites intact.See, e.g., Frommer, M., et al., Proc. Nat'l Acad. Sci. USA 1992,89:1827-31. Commercially available kits may be used for such bisulfitetreatment. In certain embodiments, to facilitate methylation PCR,primers are designed as known in the art, e.g., outer primers whichamplify DNA regardless of methylation status, and nested primers whichbind to methylated or non-methylated sequences within the regionamplified by the first PCR. See, e.g., Li et al., Bioinformatics 2002,18:1427-31. In certain embodiments, probes are designed, e.g., probeswhich bind to the bisulfite-treated DNA regardless of methylationstatus. In certain embodiments, CpG methylation is detected, e.g.,following PCR amplification of bisulfite-treated DNA using outerprimers. In certain embodiments, amplified product from the initial PCRreaction serves as a template for the nested PCR reaction usingmethylation-specific primers or non-methylation-specific primers. Incertain embodiments, a standard curve is established to determine thepercentage of methylated molecules in a particular sample. Methods fordetecting nucleic acid methylation (e.g., RNA or DNA methylation) areknown in art. See, e.g., Laird, P. W., Nature Rev. Cancer 2003,3:253-66; Belinsky, S. A., Nature Rev. Cancer 2004, 4:1-11.

In certain embodiments, statistical analyses are performed to assess theinfluence of particular methylation levels with the potential benefit oftreatment with a particular pharmaceutical composition comprising acytidine analog. In certain embodiments, the influence of methylation onoverall survival is assessed, e.g., using Cox proportional hazardsmodels and Kaplan-Meier (KM) methodology.

In certain embodiments, any gene associated with a particular solidtumor, leukemia, lymphoma, multiple myeloma, MDS, or AML may be examinedfor its methylation status in a patient. Particular genes associatedwith a solid tumor, leukemia, lymphoma, multiple myeloma, MDS, or AML,which would be suitable for use in the methods disclosed here, may beknown in the art.

In specific embodiments, provided herein is a method of identifying asubject who is likely to be responsive to a treatment described herein,comprising: (a) determining the level of a biomarker in a biologicalsample from the subject, wherein the biomarker is described herein; and(b) comparing the level of the biomarker in the biological sample to areference level of the biomarker; wherein the subject is likely to beresponsive to the treatment if the level of the biomarker in thebiological sample from the subject is altered (e.g., high or low) ascompared to the reference level of the biomarker.

In specific embodiments, provided herein is a method of identifying asubject who is likely to be responsive to a treatment described herein,comprising: (a) determining the level of a biomarker in a biologicalsample from the subject, wherein the biomarker is described herein; (b)determining the level of the biomarker in a control sample; and (c)comparing the level of the biomarker in the biological sample from thesubject to the level of the biomarker in the control sample; wherein thesubject is likely to be responsive to the treatment if the level of thebiomarker in the biological sample from the subject is altered (e.g.,high or low) as compared to the level of the biomarker in the controlsample.

In specific embodiments, provided herein is a method of identifying asubject who is likely to be responsive to a treatment described herein,comprising: (a) obtaining a biological sample from the subject; (b)determining the level of a biomarker in the biological sample, whereinthe biomarker is described herein; and (c) comparing the level of thebiomarker in the biological sample to a reference level of thebiomarker; wherein the subject is likely to be responsive to thetreatment if the level of the biomarker in the biological sample fromthe subject is altered (e.g., high or low) as compared to the referencelevel of the biomarker.

In specific embodiments, provided herein is a method of identifying asubject who is likely to be responsive to a treatment described herein,comprising: (a) obtaining a biological sample from the subject; (b)determining the level of a biomarker in the biological sample, whereinthe biomarker is described herein; (c) determining the level of thebiomarker in a control sample; and (d) comparing the level of thebiomarker in the biological sample from the subject to the level of thebiomarker in the control sample; wherein the subject is likely to beresponsive to the treatment if the level of the biomarker in thebiological sample from the subject is altered (e.g., high or low) ascompared to the level of the biomarker in the control sample.

In specific embodiments, provided herein is a method of predicting theresponsiveness of a subject to a treatment described herein, comprising:(a) determining the level of a biomarker in a biological sample from thesubject, wherein the biomarker is described herein; and (b) comparingthe level of the biomarker in the biological sample to a reference levelof the biomarker; wherein the difference between the level of thebiomarker in the biological sample from the subject and the referencelevel of the biomarker (e.g., higher or lower) correlates with theresponsiveness of the subject to the treatment.

In specific embodiments, provided herein is a method of predicting theresponsiveness of a subject to a treatment described herein, comprising:(a) determining the level of a biomarker in a biological sample from thesubject, wherein the biomarker is described herein; (b) determining thelevel of the biomarker in a control sample; and (c) comparing the levelof the biomarker in the biological sample from the subject to the levelof the biomarker in the control sample; wherein the difference betweenthe level of the biomarker in the biological sample from the subject andthe level of the biomarker in the control sample (e.g., higher or lower)correlates with the responsiveness of the subject to the treatment.

In specific embodiments, provided herein is a method of predicting theresponsiveness of a subject to a treatment described herein, comprising:(a) obtaining a biological sample from the subject; (b) determining thelevel of a biomarker in the biological sample, wherein the biomarker isdescribed herein; and (c) comparing the level of the biomarker in thebiological sample to a reference level of the biomarker; wherein thedifference between the level of the biomarker in the biological samplefrom the subject and the reference level of the biomarker (e.g., higheror lower) correlates with the responsiveness of the subject to thetreatment.

In specific embodiments, provided herein is a method of predicting theresponsiveness of a subject to a treatment described herein, comprising:(a) obtaining a biological sample from the subject; (b) determining thelevel of a biomarker in the biological sample, wherein the biomarker isdescribed herein; (c) determining the level of the biomarker in acontrol sample; and (d) comparing the level of the biomarker in thebiological sample from the subject to the level of the biomarker in thecontrol sample; wherein the difference between the level of thebiomarker in the biological sample from the subject and the level of thebiomarker in the control sample (e.g., higher or lower) correlates withthe responsiveness of the subject to the treatment.

In specific embodiments, provided herein is a method of monitoring theefficacy of a treatment described herein, comprising: (a) obtaining afirst biological sample from the subject; (b) determining the level of abiomarker in the first biological sample, wherein the biomarker isdescribed herein; (c) administering the treatment compound to thesubject; (d) thereafter obtaining a second biological sample from thesubject; (e) determining the level of the biomarker in the secondbiological sample; and (f) comparing the levels of the biomarker in thefirst and second biological samples; wherein the subject is responsiveto the treatment if the level of the biomarker in the second biologicalsample of the subject is altered (e.g., high or low) as compared to thelevel of the biomarker in the first biological sample of the subject.

In specific embodiments, provided herein is a method of monitoring thecompliance of a subject with a treatment described herein, comprising:(a) obtaining a biological sample from the subject; (b) determining thelevel of a biomarker in the biological sample, wherein the biomarker isdescribed herein; and (c) comparing the level of the biomarker with thelevel of the biomarker in a control sample from the subject; wherein thechange in the level of the biomarker in the biological sample incomparison with the level of the biomarker in the control sample (e.g.,high or low) indicates the compliance of the subject with the treatment.

3. Co-Administered Therapeutic Agents

In one embodiments, methods provided herein for treating cancer compriseco-administering a cytidine analog, such as, for example, 5-azacytidine,with one or more therapeutic agents, such as, for example, cancertherapeutic agents, to yield a synergistic therapeutic effect. In oneembodiment, the co-administered therapeutic agent is provided hereinabove (e.g., one or more of the additional therapeutic agent describedherein). In exemplary embodiments, the co-administered therapeuticagents include, but are not limited to, e.g., cytotoxic agents,anti-metabolites, antifolates, DNA intercalating agents, DNAcross-linking agents, DNA alkylating agents, DNA cleaving agents,topoisomerase inhibitors, HDAC inhibitors such as MGCD0103 (a.k.a.N-(2-aminophenyl)-4-((4-(pyridin-3-yl)pyrimidin-2-ylamino)methyl)benzamide),CDK inhibitors, JAK inhibitors, anti-angiogenic agents, Bcr-Ablinhibitors, HER2 inhibitors, EGFR inhibitors, VEGFR inhibitors, PDGFRinhibitors, HGFR inhibitors, IGFR inhibitors, c-Kit inhibitors, Raspathway inhibitors, PI3K inhibitors, multi-targeted kinase inhibitors,mTOR inhibitors, anti-estrogens, anti-androgens, aromatase inhibitors,somatostatin analogs, ER modulators, anti-tubulin agents, vincaalkaloids, taxanes, HSP inhibitors, Smoothened antagonists, telomeraseinhibitors, COX-2 inhibitors, anti-metastatic agents,immunosuppressants, biologics such as antibodies, and hormonaltherapies. In particular embodiment, the co-administered therapeuticagent is thalidomide, lenalidomide, or pomalidomide. In particularembodiment, the co-administered therapeutic agent is carboplatin. Inparticular embodiment, the co-administered therapeutic agent ispaclitaxel (e.g., Abraxane®). See, e.g., U.S. Pat. Nos. 7,758,891,7,771,751, 7,820,788, 7,923,536, 8,034,375; U.S. Patent Publication No.2010/0048499; all of which are incorporated herein by reference in theirentireties. The co-administered agent may be dosed, e.g., orally or byinjection.

In one embodiment, the route of the administration of the cytidineanalog (e.g., 5-azacytidine or another cytidine analog provided herein)is independent of the route of the administration of a second therapy.In one embodiment, the cytidine analog is administered orally. Inanother embodiment, the cytidine analog is administered intravenously orsubcutaneously. In certain embodiments, the cytidine analog isadministered orally, and the second therapy is administered orally,parenterally, intraperitoneally, intravenously, intraarterially,transdermally, sublingually, intramuscularly, rectally, transbuccally,intranasally, liposomally, via inhalation, vaginally, intraoccularly,via local delivery by catheter or stent, subcutaneously,intraadiposally, intraarticularly, intrathecally, or in a slow releasedosage form. In one embodiment, the cytidine analog and a second therapyare administered by the same mode of administration, e.g., orally,intravenously, or subcutaneously. In another embodiment, the cytidineanalog is administered by one mode of administration, e.g., orally,whereas the second agent (e.g., an anticancer agent) is administered byanother mode of administration, e.g., intravenously or subcutaneously.In yet another embodiment, the cytidine analog is administered by onemode of administration, e.g., intravenously or subcutaneously, whereasthe second agent (e.g., an anticancer agent) is administered by anothermode of administration, e.g., orally.

In one embodiment, each method provided herein may independently,further comprise the step of administering a second therapeutic agent.In one embodiment, the second therapeutic agent is an anticancer agent.In one embodiment, the anticancer agent is an antimetabolite, including,but not limited to, 5-fluoro uracil, methotrexate, cytarabine, high dosecytarabine, and fludarabine. In one embodiment, the anticancer agent isan antimicrotubule agent, including, but not limited to, vinca alkaloids(e.g., vincristine and vinblastine) and taxanes (e.g., paclitaxel, e.g.,Abraxane®, and docetaxel). In one embodiment, the anticancer agent is analkylating agent, including, but not limited to, cyclophosphamide,melphalan, carmustine, and nitrosoureas (e.g., hydroxyurea andbischloroethylnitrosurea). In one embodiment, the anticancer agent is aplatinum agent, including, but not limited to, cisplatin, carboplatin,oxaliplatin, satraplatin (JM-216), and CI-973. In one embodiment, theanticancer agent is an anthracycline, including, but not limited to,doxrubicin and daunorubicin. In one embodiment, the anticancer agent isan antitumor antibiotic, including, but not limited to, mitomycin,idarubicin, adriamycin, and daunomycin (also known as daunorubicin). Inone embodiment, the anticancer agent is a topoisomerase inhibitor, e.g.,etoposide and camptothecins. In one embodiment, the anticancer agent isselected from the group consisting of adriamycin, busulfan, cytarabine,cyclophosphamide, dexamethasone, fludarabine, fluorouracil, hydroxyurea,interferons, oblimersen, platinum derivatives, taxol, topotecan, andvincristine.

In one embodiment, other therapies or anticancer agents that may be usedin combination with the cytidine analog include surgery, radiotherapy(e.g., gamma-radiation, neutron beam radiotherapy, electron beamradiotherapy, proton therapy, brachytherapy, and systemic radioactiveisotopes), endocrine therapy, biologic response modifiers (e.g.,interferons, interleukins, and tumor necrosis factor (TNF)),hyperthermia and cryotherapy, agents to attenuate any adverse effects(e.g., antiemetics), and other approved chemotherapeutic drugs,including, but not limited to, alkylating drugs (mechlorethamine,chlorambucil, cyclophosphamide, melphalan, and ifosfamide),antimetabolites (cytarabine, high dose cytarabine, and methotrexate),purine antagonists and pyrimidine antagonists (6-mercaptopurine,5-fluorouracil, cytarabine, and gemcitabine), spindle poisons(vinblastine, vincristine, vinorelbine, docetaxel, and paclitaxel, e.g.,Abraxane®), podophyllotoxins (etoposide, irinotecan, and topotecan),antibiotics (daunorubicin, doxorubicin, bleomycin, and mitomycin),nitrosoureas (carmustine and lomustine), inorganic ions (cisplatin andcarboplatin), enzymes (asparaginase), and hormones (tamoxifen,leuprolide, flutamide, and megestrol), imatinib, adriamycin,dexamethasone, and cyclophosphamide. For additional available cancertherapies, see, e.g., http://www.nci.nih.gov/; for a list of FDAapproved oncology drugs, see, e.g., http://www.fda.gov/, The MerckManual, 18th Ed. 2006, and PDR: Physician Desk Reference 2010, 64th Ed.2009; the contents of each of which are hereby incorporated by referencein their entireties.

In one embodiment, without being limited by a particular theory,methylation-based silencing of specific genes limits the anti-tumoreffects of cytotoxic agents. In one embodiment, without being limited bya particular theory, a cytidine analog, such as, for example,5-azacytidine or decitabine, can sensitize tumors to the effects ofchemotherapy (e.g., the effect of an anti-cancer agent). In oneembodiment, without being limited by a particular theory, the epigeneticeffect of a cytidine analog, such as, for example, 5-azacytidine ordecitabine, restores chemo-sensitivity of cancer cells, after the cancercells are contacted with the cytidine analog for a period of time. Incertain embodiments, without being limited by a particular theory, acytidine analog is administered to a subject in need thereof for asustained period of time (e.g., multiple doses or multiple treatmentcycles) before the subject is treated with an additional therapeuticagent (e.g., an anti-cancer agent) to yield a greater synergistictherapeutic effect and/or a reduced toxicity effect. In someembodiments, without being limited by a particular theory,co-administration of a cytidine analog and certain anti-cancer agent(e.g., a cytotoxic agent) from the first day of therapy may produceincreased toxicity without added anti-tumor effects. In someembodiments, without being limited by a particular theory, sustainedexposure of a subject to a cytidine analog (e.g., 5-azacytidine ordecitabine or another cytidine analog provided herein) prior to theadministration of an additional therapeutic agent (e.g., a cytotoxicagent) yield a synergistic therapeutic effect (e.g., sensitization ofcancer cells to the cytotoxic agent).

In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 100 mg/day for 7 days or morebefore a second therapeutic agent is administered to the subject. Inparticular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 100 mg/day for 14 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 100 mg/day for 21 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 100 mg/day for 28 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 150 mg/day for 7 days or morebefore a second therapeutic agent is administered to the subject. Inparticular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 150 mg/day for 14 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 150 mg/day for 21 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 150 mg/day for 28 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 200 mg/day for 7 days or morebefore a second therapeutic agent is administered to the subject. Inparticular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 200 mg/day for 14 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 200 mg/day for 21 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 200 mg/day for 28 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 250 mg/day for 7 days or morebefore a second therapeutic agent is administered to the subject. Inparticular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 250 mg/day for 14 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 250 mg/day for 21 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 250 mg/day for 28 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 300 mg/day for 7 days or morebefore a second therapeutic agent is administered to the subject. Inparticular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 300 mg/day for 14 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 300 mg/day for 21 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 300 mg/day for 28 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 350 mg/day for 7 days or morebefore a second therapeutic agent is administered to the subject. Inparticular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 350 mg/day for 14 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 350 mg/day for 21 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 350 mg/day for 28 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 400 mg/day for 7 days or morebefore a second therapeutic agent is administered to the subject. Inparticular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 400 mg/day for 14 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 400 mg/day for 21 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 400 mg/day for 28 days ormore before a second therapeutic agent is administered to the subject.In particular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 450 mg/day for 7 days or morebefore a second therapeutic agent is administered to the subject. Inparticular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 480 mg/day for 7 days or morebefore a second therapeutic agent is administered to the subject. Inparticular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 500 mg/day for 7 days or morebefore a second therapeutic agent is administered to the subject. Inparticular embodiments, 5-azacytidine is administered orally to asubject in need thereof at a dose of about 600 mg/day for 7 days or morebefore a second therapeutic agent is administered to the subject.

In one embodiment, after the second therapeutic agent is administered,the administration of the cytidine analog is continued for 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 14, 16, 18, 20, 22, 24, 26, 28, or more than28 days; optionally followed with a resting period from theadministration of the cytidine analog of about 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 12, 14, 21, 28, or more than 28 days.

In one embodiment, the second therapeutic agent is administeredcyclically, after the first dose. In one embodiment, the methodsprovided herein comprise: i) administering to the subject a first dailydose of the second therapeutic agent; ii) optionally resting for aperiod of at least one day where the second therapeutic agent is notadministered to the subject; iii) administering a second dose of thesecond therapeutic agent to the subject; and iv) repeating steps ii) toiii) a plurality of times. In certain embodiments, the first daily doseis between about 50 mg/m²/day and about 2,000 mg/m²/day. In certainembodiments, the second daily dose is between about 50 mg/m²/day andabout 2,000 mg/m²/day. In certain embodiments, the first daily dose isbetween about 50 mg/m²/day and about 200 mg/m²/day. In certainembodiments, the second daily dose is between about 50 mg/m²/day andabout 200 mg/m²/day. In certain embodiments, the first daily dose isbetween about 100 mg/day and about 1,000 mg/day. In certain embodiments,the second daily dose is between about 100 mg/day and about 1,000mg/day. In certain embodiments, the first daily dose is higher than thesecond daily dose. In certain embodiments, the second daily dose ishigher than the first daily dose. In certain embodiments, the seconddaily dose and the first daily dose are the same. In one embodiment, therest period is 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 15 days, 16days, 17 days, 18 days, 19 days, 20 days, 21 days, or 28 days. In oneembodiment, the rest period is at least 2 days and steps ii) throughiii) are repeated at least three times. In one embodiment, the restperiod is at least 2 days and steps ii) through iii) are repeated atleast five times. In one embodiment, the rest period is at least 3 daysand steps ii) through iii) are repeated at least three times. In oneembodiment, the rest period is at least 3 days and steps ii) throughiii) are repeated at least five times. In one embodiment, the restperiod is at least 7 days and steps ii) through iii) are repeated atleast three times. In one embodiment, the rest period is at least 7 daysand steps ii) through iii) are repeated at least five times. In oneembodiment, the rest period is at least 14 days and steps ii) throughiii) are repeated at least three times. In one embodiment, the restperiod is at least 14 days and steps ii) through iii) are repeated atleast five times. In one embodiment, the rest period is at least 21 daysand steps ii) through iii) are repeated at least three times. In oneembodiment, the rest period is at least 21 days and steps ii) throughiii) are repeated at least five times. In one embodiment, the restperiod is at least 28 days and steps ii) through iii) are repeated atleast three times. In one embodiment, the rest period is at least 28days and steps ii) through iii) are repeated at least five times.

In one embodiment, 5-azacytidine is administered orally for 7 days outof a 28-day cycle. In one embodiment, 5-azacytidine is administeredorally for 14 days out of a 28-day cycle. In one embodiment,5-azacytidine is administered orally for 21 days out of a 28-day cycle.In one embodiment, 5-azacytidine is administered orally for 7 days outof a 21-day cycle. In one embodiment, 5-azacytidine is administeredorally for 14 days out of a 21-day cycle. In one embodiment,5-azacytidine is administered orally for 21 days out of a 21-day cycle.In one embodiment, 5-azacytidine is administered orally once daily. Inone embodiment, 5-azacytidine is administered orally twice daily. In oneembodiment, 5-azacytidine is administered orally once daily in an amountof about 50 mg/day. In one embodiment, 5-azacytidine is administeredorally twice daily in an amount of about 50 mg/day. In one embodiment,5-azacytidine is administered orally once daily in an amount of about 50mg/day for 7, 14, or 21 days. In one embodiment, 5-azacytidine isadministered orally twice daily in an amount of about 50 mg/day for 7,14, or 21 days. In one embodiment, 5-azacytidine is administered orallyonce daily in an amount of about 50 mg/day for more than 21 days. In oneembodiment, 5-azacytidine is administered orally once daily in an amountof about 100 mg/day. In one embodiment, 5-azacytidine is administeredorally twice daily in an amount of about 100 mg/day. In one embodiment,5-azacytidine is administered orally once daily in an amount of about100 mg/day for 7, 14, or 21 days. In one embodiment, 5-azacytidine isadministered orally twice daily in an amount of about 100 mg/day for 7,14, or 21 days. In one embodiment, 5-azacytidine is administered orallyonce daily in an amount of about 100 mg/day for more than 21 days. Inone embodiment, 5-azacytidine is administered orally once daily in anamount of about 150 mg/day. In one embodiment, 5-azacytidine isadministered orally twice daily in an amount of about 150 mg/day. In oneembodiment, 5-azacytidine is administered orally once daily in an amountof about 150 mg/day for 7, 14, or 21 days. In one embodiment,5-azacytidine is administered orally twice daily in an amount of about150 mg/day for 7, 14, or 21 days. In one embodiment, 5-azacytidine isadministered orally once daily in an amount of about 150 mg/day for morethan 21 days. In one embodiment, 5-azacytidine is administered orallyonce daily in an amount of about 200 mg/day. In one embodiment,5-azacytidine is administered orally twice daily in an amount of about200 mg/day. In one embodiment, 5-azacytidine is administered orally oncedaily in an amount of about 200 mg/day for 7, 14, or 21 days. In oneembodiment, 5-azacytidine is administered orally twice daily in anamount of about 200 mg/day for 7, 14, or 21 days. In one embodiment,5-azacytidine is administered orally once daily in an amount of about200 mg/day for more than 21 days. In one embodiment, 5-azacytidine isadministered orally once daily in an amount of about 250 mg/day. In oneembodiment, 5-azacytidine is administered orally twice daily in anamount of about 250 mg/day. In one embodiment, 5-azacytidine isadministered orally once daily in an amount of about 250 mg/day for 7,14, or 21 days. In one embodiment, 5-azacytidine is administered orallytwice daily in an amount of about 250 mg/day for 7, 14, or 21 days. Inone embodiment, 5-azacytidine is administered orally once daily in anamount of about 250 mg/day for more than 21 days. In one embodiment,5-azacytidine is administered orally once daily in an amount of about300 mg/day. In one embodiment, 5-azacytidine is administered orallytwice daily in an amount of about 300 mg/day. In one embodiment,5-azacytidine is administered orally once daily in an amount of about300 mg/day for 7, 14, or 21 days. In one embodiment, 5-azacytidine isadministered orally twice daily in an amount of about 300 mg/day for 7,14, or 21 days. In one embodiment, 5-azacytidine is administered orallyonce daily in an amount of about 300 mg/day for more than 21 days. Inparticular embodiments, 5-azacytidine is administered continuously for14 days, followed with a 7-day resting period.

4. Methods Comprising Co-Administering One or More AdditionalTherapeutic Agents with Oral Formulations Disclosed Herein

Particular embodiments herein provide methods of treating diseases ordisorders disclosed herein (e.g., diseases or disorders involvingabnormal cell proliferation), wherein the methods compriseco-administering an oral formulation disclosed herein, such as, forexample, an oral formulation comprising 5-azacytidine or anothercytidine analog provided herein, with one or more additional therapeuticagents (such as, for example, a cancer therapeutic agent) to yield asynergistic therapeutic effect. Particular co-administered therapeuticagents useful in the methods disclosed herein are disclosed throughoutthe specification. In particular embodiments, the co-administeredtherapeutic agent is carboplatin. In particular embodiments, theco-administered therapeutic agent is paclitaxel (e.g., Abraxane®). Inparticular embodiments, the additional therapeutic agent isco-administered in an amount that is a therapeutically effective amount.In particular embodiments, the additional therapeutic agent isco-administered in a separate dosage form from the cytidine analogdosage form with which it is co-administered. In particular embodiments,the additional therapeutic agent is co-administered in a dosage form(e.g., a single unit dosage form) together with the cytidine analog withwhich it is co-administered. In such cases, the cytidine analog and theadditional therapeutic agent may be co-formulated together in the samedosage form using methods of co-formulating active pharmaceuticalingredients, including methods disclosed herein and methods known in theart.

In particular embodiments, a cytidine analog is administered to asubject in need thereof, for a sustained period of time (e.g., for 1, 2,3, 4, 5, 6, 7, or more than 7 days) before one or more additionaltherapeutic agent(s) is/are administered to the subject. In particularembodiments, provided herein are methods of treating diseases ordisorders disclosed herein (e.g., diseases or disorders involvingabnormal cell proliferation, such as a relapsed or refractory solidtumor), wherein the methods comprise: (i) first administering a cytidineanalog orally to a subject in need thereof, for 1, 2, 3, 4, 5, 6, 7, ormore than 7 days; and (ii) administering an additional therapeutic agent(e.g., an anti-cancer agent provided herein, such as, carboplatin orpaclitaxel, e.g., Abraxane®) for one or more days. In certainembodiment, the second step comprises continued administration of thecytidine analog orally for one or more additional days (e.g., for 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or more than 14 days).

INCORPORATION BY REFERENCE

All disclosures (e.g., patents, publications, and web pages) referencedthroughout this specification are incorporated by reference in theirentireties.

V. EXAMPLES A. Example 1

Clinical studies were conducted to evaluate oral azacitidine as a singleagent and in combination with Carboplatin or Abraxane® in subjects withrelapsed or refractory solid tumors.

One objective of the study was to evaluate the safety and to define theMaximal Tolerated Dose (MTD) or the Maximal Administered Dose (MAD) oforal azacitidine as a single agent and in combination with carboplatin(CBDCA) or paclitaxel protein-bound particles (Abraxane® [ABX]) insubjects with relapsed or refractory solid tumors.

Other objectives of the study included: (1) to examine the impact, ifany, of CBDCA or ABX on the pharmacokinetics (PK) of oral azacitidine;(2) to examine the impact, if any, of oral azacitidine on the PK ofCBDCA or ABX; (3) to evaluate the pharmacodynamic (PD) effects of oralazacitidine as a single agent and in combination with CBDCA and ABX inblood, plasma and tumor tissue; and (4) to make a preliminary assessmentof the anti-tumor activity of oral azacitidine as a single agent and incombination with CBDCA and ABX in specific tumor types.

Additional objectives of the study included: to determine whether thereis any relationship among baseline tumor molecular characteristics(genetic or epigenetic), PD effects, and anti-tumor activity.

The study was an open-label, 3-arm, multi-center, dose-escalation studyof oral azacitidine in combination with either CBDCA (Arm A), ABX (ArmB), or as a single agent (Arm C) in subjects with relapsed or refractorysolid tumors (Part 1). Subjects were assigned to each study Arm at thediscretion of the investigator. A minimum of 6 subjects were assigned toeach study Arm when a dose level (DL) became open for enrollment. If one(1) or zero (0) out of six (6) subjects in a DL experienced doselimiting toxicity (DLT), the dose of oral azacitidine was escalated inthe successive DL. A limited number of oral azacitidine DLs wereexplored to arrive at a recommended Part 2 dose (RP2D) of oralazacitidine for each study Arm. The RP2D may be the MTD, MAD, or a lowerdose depending on the tolerability, PK, and PD observed. Part 1 wasfollowed by expansion cohorts at the RP2D in specific tumor types (Part2). Approximately 60 subjects were enrolled in Part 1, and approximately100 subjects were enrolled in Part 2. Safety, efficacy,pharmacokinetics, and pharmacodynamics data were evaluated.

In Part 1, Arms A and B, Cycle 1 was 28 days in duration. Subsequenttreatment Cycles were 21 days in duration. In Arm C, all Cycles were 21days in duration.

Part 1 Design:

Subjects may continue to receive their assigned combination treatment ifthey have no unacceptable toxicity and if there is no clinical orradiographic evidence of disease progression or the investigator deemsthat the subject is deriving potential benefit. If combination treatmentis suspended for unacceptable toxicity that is believed to be related toCBDCA in Arm A or ABX in Arm B, subjects may continue to take singleagent oral azacitidine at their assigned DL once the toxicity resolvesto at least grade 1. Subjects in Arm C receive single agent oralazacitidine in all Cycles up to approximately 1 year from the start oftherapy or until they experience unacceptable toxicity or progressivedisease, as assessed by the investigator, whichever occurs first.Escalation of the oral azacitidine dose continues independently in eachArm until the RP2D of oral azacitidine as a single agent and incombination with CBDCA and ABX is defined. The RP2D may be different foreach study Arm.

Part 2 Design:

Expansion cohorts of up to 20 subjects for each of several specifictumor types are enrolled at the RP2D for each Arm. In addition tofurther exploring the safety and PD activity of oral azacitidine aloneand in combination with CBDCA or ABX in specific tumor types, this partof the study is designed to make an initial assessment of anti-tumoractivity and its potential association with candidate predictivebiomarkers. Tumor biopsies are performed in Part 2.

Study Population:

Men and women, 18 years or older, with histological or cytologicalconfirmation of advanced unresectable solid tumors, including those whohave progressed on (or not been able to tolerate) standard anti-cancertherapy, or for whom no other known effective therapy exists, or forthose who have declined standard therapy.

length of study: the duration of part 1 of the study from first subjectscreened to last subject last visit is approximately 1 year. Part 2 ofthe study lasts approximately 18 months making the entire duration ofthe study approximately 2.5 years.

Study Treatments:

Part 1:

Subjects receive oral azacitidine as a single agent for the first 7 daysof study. Beginning on Cycle 1, Day 8, subjects in Arms A and B begincombination treatment with CBDCA or ABX, respectively. Subjects maycontinue to receive their assigned combination until they experiencedisease progression or unacceptable toxicity, whichever occurs first.Subjects in Arm C receive single agent oral azacitidine in all Cyclesuntil they experience unacceptable toxicity or progressive disease,whichever occurs first.

The dose of oral azacitidine in each Arm is escalated (or reduced) basedon tolerability with a fixed dose of CBDCA or ABX in the first 28-dayCycle, or as a single agent in the first 21-day Cycle, until the RP2D isdefined.

All study Arms begin at Dose Level 1 (DL1). If DL1 is declaredtolerable, Dose Level 2 (DL2) opens for enrollment. If DL2 is declaredtolerable, this dose and schedule are explored in Part 2 of the study.If DL2 exceeds the maximum tolerated dose (MTD), DL1 is explored in Part2. If DL1 exceeds MTD, Dose Level-1 (DL-1) opens for enrollment.

For Arms A and B, if DL-1 is declared tolerable, Dose Level-2 (DL-2)opens for enrollment. If DL-2 is declared tolerable, this dose andschedule are explored in Part 2. If DL-2 exceeds MTD, DL-1 is exploredin Part 2. If DL-1 exceeds MTD, enrollment to that Arm stops (FIG. 4).

For Arm C, if DL-1 is declared tolerable, that dose and schedule areexplored in Part 2. If DL-1 exceeds MTD, DL-2 opens for enrollment. IfDL-2 is declared tolerable, that dose and schedule are explored in Part2. If DL-2 exceeds MTD, enrollment into Arm C stops (FIG. 5).

In certain embodiments, subjects receive a dose of a prophylacticanti-emetic, for example, a 5-HT3 antagonist, prior to each dose of oralazacitidine.

Part 2:

Subjects in Part 2 of the study receive oral azacitidine alone (Arm C)or in combination with CBDCA (Arm A) or ABX (Arm B) according to theRP2D established for each Arm in Part 1. All treatment Cycles in Part 2are 21 days in duration. Each specific tumor type in Part 2 receivestreatment according to one of the three Arms. About 14 to 20 patientsare enrolled per tumor type.

Assignment of Subject to Study Arms:

At the time of enrollment in Part 1, subjects who meet all of theinclusion criteria and none of the exclusion criteria are assigned toeither Arm A (oral azacitidine with CBDCA), Arm B (oral azacitidine withAbraxane®), or Arm C (single agent oral azacitidine). Assignment ofsubjects to each of these study Arms is at the discretion of theinvestigator, based on the suitability of the regimen(s) for the subjectand availability of open enrollment slots. The DLs for Arms A, B, and Care shown in Table 1, Table 2, and Table 3, respectively.

TABLE 1 Oral Azacitidine and Carboplatin (CBDCA) Dose Levels for Arm ACycle 1 (28 days) Cycle 2 (21 days) Cycles 3+ (21 days) Oral Oral OralAzacitidine CBDCA Azacitidine CBCDA Azacitidine CBCDA DL-2 100 mg AUC4100 mg AUC 4 100 mg AUC 4 Days 1-7, 9-28 Day 8 Days 1-21 Day 1 Days 1-21Day 1 DL-1 100 mg AUC4 100 mg AUC 4 100 mg AUC 4 Days 1-7, 9-14, Day 8Days 1-7, Day 1 Days 1-7, Day 1 22-28 15-21 15-21 DL1 200 mg AUC4 200 mgAUC 4 200 mg AUC 4 Days 1-7, 9-14, Day 8 Days 1-7, Day 1 Days 1-7, Day 122-28 15-21 15-21 DL2 300 mg AUC4 300 mg AUC 4 300 mg AUC 4 Days 1-7,9-14, Day 8 Days 1-7, Day 1 Days 1-7, Day 1 22-28 15-21 15-21

TABLE 2 Oral Azacitidine and Abraxane ® (ABX) Dose Levels for Arm BCycle 1 (28 days) Cycle 2 (21 days) Cycles 3+ (21 days) Oral Oral OralAzacitidine ABX Azacitidine ABX Azacitidine ABX DL-2 100 mg 100 mg/m²100 mg 100 mg/m² 100 mg 100 mg/m² Days 1-14, Days 8, 15 Days 1-21 Days1, 8 Days 1-21 Days 1, 8 19-28 DL-1 100 mg 100 mg/m² 100 mg 100 mg/m²100 mg 100 mg/m² Days 1-14, Days 8, 15 Days 1-7, Days 1, 8 Days 1-7,Days 1, 8 22-28 15-21 15-21 DL1 200 mg 100 mg/m² 200 mg 100 mg/m² 200 mg100 mg/m² Days 1-14, Days 8, 15 Days 1-7, Days 1, 8 Days 1-7, Days 1, 822-28 15-21 15-21 DL2 300 mg 100 mg/m² 300 mg 100 mg/m² 300 mg 100 mg/m²Days 1-14, Days 8, 15 Days 1-7, Days 1, 8 Days 1-7, Days 1, 8 22-2815-21 15-21

TABLE 3 Oral Azacitidine Dose Levels for Arm C Cycle 1 Cycle 2 Cycles 3+(21 days) (21 days) (21 days) Oral Azacitidine DL-2 200 mg, Days 1-14200 mg, Days 1-14 200 mg, Days 1-14 DL-1 300 mg, Days 1-14 300 mg, Days1-14 300 mg, Days 1-14 DL1 200 mg, Days 1-21 200 mg, Days 1-21 200 mg.Days 1-21 DL2 300 mg, Days 1-21 300 mg, Days 1-21 300 mg. Days 1-21

Efficacy Assessments:

Subjects are evaluated for tumor response after Cycle 2 and every otherCycle thereafter. The primary efficacy variables are tumor response atthe end of treatment, and the proportion of subjects alive andprogression-free (progression-free survival; PFS) at the end of Cycle 6.Tumor response is based on Response Evaluation Criteria in Solid Tumors(RECIST) 1.1 for disease states which require at least one measurabletarget lesion at baseline for study eligibility. Progression-freesurvival rates are computed using the Kaplan-Meier estimates. Durationof response is reported in subjects who have a complete or partialresponse. Ninety percent confidence intervals (90% CIs) of the responserate at the end of treatment, and of the PFS rate at time of eachscheduled response assessment (i.e., Cycles 2, 4, 6, etc.) are providedby tumor type. Other endpoints that are explored includetime-to-tumor-progression and overall survival.

The influence of major disease characteristics and prognosticindications are considered in relationship to efficacy, with specialattention given to subjects in Arm A who were previously treated with aplatin and to subjects in Arm B who were previously treated with ataxane. Full details on the efficacy analysis are given in theStatistical Analysis Plan (SAP).

Safety Assessments:

Safety assessments include adverse events (AEs), physical examinations(PEs), (including height and body weight); vital signs (includingsystolic/diastolic blood pressure [BP], pulse rate, respiratory rate,and oral temperature); Eastern Cooperative Oncology Group (ECOG)performance status; 12-lead electrocardiogram (ECG [including rhythm,heart rate, PR, QRS, and QT intervals]); complete blood count (CBC)(including hemoglobin, hematocrit, red blood cell count with indices[mean corpuscular volume {MCV}, mean corpuscular hemoglobin {MCH}, andmean corpuscular hemoglobin concentration {MCHC}, white blood cell [WBC]count with absolute differential [neutrophils, lymphocytes, monocytes,eosinophils, and basophils], and platelet count); coagulation(international normalized ratio [INR], prothrombin time [PT], andpartial thromboplastin time [PTT]); serum chemistry (non-fasting)(including albumin, alkaline phosphatase, bicarbonate, blood ureanitrogen [BUN], calcium, chloride, creatinine, glucose, lacticdehydrogenase [LDH], phosphorus, potassium, aspartate aminotransferase[AST], alanine aminotransferase [ALT], sodium, total bilirubin, totalprotein, and uric acid); screening serum pregnancy test required for allfemales of child-bearing potential (FCBP) prior to Cycle 1 Day 1 dosing(test result must be obtained and read prior to dosing on Day 1.); tumorbiopsy (optional); and tumor assessment.

Study Endpoints:

The nature, incidence and severity of AEs are evaluated using theNational Cancer Institute Common Terminology Criteria for Adverse Events(NCI CTCAE) criteria, Version 4.0. For oral azacitidine, CBDCA, and ABX(administered alone and in combination), the following plasma PKparameters are assessed: (1) maximum observed concentration in plasma(C_(max)); (2) area under the concentration-time curve (AUC); (3) timeto maximum concentration (t_(max)); (4) terminal half-life (t₁₁₂); (5)apparent total body clearance (CL/F); and (6) apparent volume ofdistribution (Vz/F). To evaluate the PD effects of oral azacitidine inblood, plasma, and tumor tissue, the following endpoints are collected:(1) change from baseline (Cycle 1 Day 1 pre-dose) in DNA methylation(global and gene-specific assays) in whole blood and tumor tissue (asavailable in Part 1); (2) reduction from baseline (Cycle 1 Day 1predose) in DNMT1 protein levels in tumor tissue (as available in Part1). Anti-tumor activity endpoints using tumor-specific response criteriafor each tumor type include: (1) response rate and duration of response;and (2) progression-free survival (PFS). Molecular characteristics ofthe blood and tumor, including, but not limited to, DNA/RNA methylation,gene sequence and mRNA/miRNA expression, may be evaluated at baselineand post-therapy for examination in relation to tumor responses.

Part 1/Arm A:

Subjects in Arm A receive their first dose of oral azacitidine at theirassigned DL in the clinic on Cycle 1 Day 1 along with PK (predosethrough 8 hours post-dose oral azacitidine) and predose PD blood(mandatory) and tumor (optional) sampling. On Days 2 through 7, subjectsself-administer oral azacitidine daily according to their assigned DL.Subjects return to the clinic for CBDCA dosing on Day 8 along withpredose PD sampling (blood). CBDCA is administered at AUC=4 using theGlomerular Filtration Rate (GFR) calculation from the Modification ofDiet in Renal Disease (MDRD) formula below as an i.v. infusion over 1hour.

Modification of Diet in Renal Disease (MDRD) Equation for GFR: ThisIDMS-traceable MDRD study equation calculator is for use with Scrreported in mg/dL:

GFR (mL/min/1.73 m²)=175×(S _(cr))^(−1.154)×(Age)^(−0.203)×(0.742 iffemale)×(1.212 if African American) (conventional units)

The equation does not require weight because the results are reportednormalized to 1.73 m² body surface area (BSA), which is an acceptedaverage adult surface area.

CBDCA dose (mg)=4 (GFR+25)

An on-line calculator can be found at the following link:

http://www.nkdep.nih.gov/professionals/gfr_calculators/idms_con.htm

In some embodiments, oral azacitidine is not administered on Day 8 sothat the PK profile of CBDCA alone can be established. On Day 9,subjects return to the clinic for CBDCA PK sampling approximately 24hours after the end of the initial infusion and before administration oforal azacitidine. On Day 9, subjects receive their dose of oralazacitidine at their assigned DL in the clinic. On Days 10 through 14,subjects self-administer oral azacitidine. Subjects return to the clinicon Cycle 1 Day 15 for blood PD sampling (mandatory) and tumor biopsy(optional). On Days 15 through 21 no study medication is administered(except for subjects in DL-2, who self-administer oral azacitidinedaily). On Day 22, subjects return to clinic for administration of oralazacitidine with predose blood collection (mandatory) for PD analysis.On Days 23 through 28 of Cycle 1, subjects self-administer oralazacitidine daily according to their assigned DL (FIG. 1).

Subjects who complete Cycle 1 meet the following hematologic criteria atthe beginning of each subsequent Cycle: (1) ANC>1.5×10⁹ L; and (2)Platelets>75×10⁹/L.

If the hematologic criteria are not met, the start of Cycle 2 may bedelayed for up to 7 days to allow the counts to recover. If recovery hasnot occurred after 7 days, this is considered a DLT.

In Cycle 2, Arm A subjects receive oral azacitidine in the clinic on Day1 followed by CBDCA AUC=4 as an i.v. infusion over 1 hour, along with PK(predose through 8 hours following the end of the CBDCA infusion). OnDay 2, subjects return to the clinic for PK sampling approximately 24hours following the end of the CBDCA infusion. On Days 2 through 7,subjects self-administer oral azacitidine daily according to theirassigned DL. On Days 8 through 14 of Cycle 2, no study medication isadministered (except for subjects in DL-2 who self-administer oralazacitidine at their assigned DL). On Days 15 through 21, subjectsself-administer oral azacitidine at their assigned DL.

In Cycles 3 and beyond, Arm A subjects receive oral azacitidine at theirassigned DL in the clinic on Day 1 followed by CBDCA AUC=4 as an i.v.infusion over 1 hour. On Days 2 through 7, subjects self-administer oralazacitidine daily according to their assigned DL. On Days 8 through 14,no study medication is administered (except for subjects in DL-2 whoself-administer oral azacitidine at their assigned DL). On Days 15through 21, subjects self-administer oral azacitidine at their assignedDL. Subjects may continue to receive their assigned combinationtreatment if they have no unacceptable toxicity and if there is noclinical or radiographic evidence of disease progression or they arederiving potential benefit as assessed by the investigator. Ifcombination treatment is suspended for unacceptable toxicity that isbelieved to be related to CBDCA, subjects may continue to take singleagent oral azacitidine at their assigned DL once the toxicity resolves.

Part 1/Arm B:

Subjects in Arm B receive their first dose of oral azacitidine at theirassigned DL in the clinic on Cycle 1 Day 1 along with PK (predosethrough 8 hours post-dose oral azacitidine) and PD blood sampling. Tumorbiopsy (optional) is obtained prior to the first dose of oralazacitidine on Day 1. On Days 2 through 7, subjects self-administer oralazacitidine daily according to their assigned DL. Subjects return to theclinic on Day 8 for oral azacitidine followed by ABX 100 mg/m² i.v.,along with PK (predose through approximately 8 hours post end of ABXinfusion) and predose PD blood sampling. Subjects return to the clinicon Days 9, 10 and 11 for ABX PK sampling approximately 24, 48 and 72hours from the end of the ABX infusion. On Days 9 through 14, subjectsself-administer oral azacitidine at their assigned DL. On Cycle 1 Day15, subjects report to the clinic for blood PD sampling (mandatory) andtumor biopsy (optional). Abraxane® 100 mg/m² i.v is administered onCycle 1 Day 15 followed by PK sampling (predose through approximately 8hours after the end of the ABX infusion). Subjects return to the clinicon Days 16, 17, and 18 for ABX PK sampling approximately 24, 48 and 72hours from the end of the ABX infusion. On Days 15 through 21, no oralazacitidine is administered (except for subjects assigned to DL-2 whoself-administer oral azacitidine daily according to their assigned DL onDays 19 through 21). In some embodiments, oral azacitidine is notadministered on Days 15 through 18 of Cycle 1 for subjects in DL-2 sothat the PK profile of ABX alone can be established. On Day 22, subjectsreturn to the clinic for oral azacitidine followed by ABX 100 mg/m² i.v.in the clinic after obtaining predose blood (mandatory) PD sampling. OnDays 23 through 28, subjects self-administer oral azacitidine dailyaccording to their assigned DL (FIG. 2).

Subjects who complete Cycle 1 meet the following hematologic criteria atthe beginning of each subsequent Cycle: (1) Absolute Neutrophil Count(ANC)>1.5×10⁹/L; and (2) Platelets>75×10⁹/L.

If the hematologic criteria are not met, the start of Cycle 2 may bedelayed for up to 7 days to allow the hematologic counts to recover. Ifrecovery has not occurred after 7 days, this is considered a DLT.

In Cycle 2, Arm B subjects receive oral azacitidine followed by ABX 100mg/m² i.v on Day 1. On Days 2 through 7, subjects self-administer oralazacitidine at their assigned DL. Subjects return to the clinic on Day 8for ABX 100 mg/m² i.v. In some embodiments, oral azacitidine is notadministered on Days 8 through 14 (except for subjects in DL-2 whoself-administer oral azacitidine at their assigned DL). Oral azacitidinefollowed by ABX 100 mg/m² i.v. is administered on Day 15. On Days 16through 21, subjects self-administer oral azacitidine at their assignedDL. Subjects who complete Cycle 2 without unacceptable toxicity andwithout objective evidence of disease progression as per a tumorassessment may proceed to Cycle 3.

In Cycle 3 and beyond, Arm B subjects receive oral azacitidine at theirassigned DL in the clinic on Day 1, followed by ABX 100 mg/m² i.v. OnDays 2 through 7, subjects self-administer oral azacitidine at theirassigned DL. Subjects return to the clinic on Day 8 for ABX 100 mg/m²i.v. In some embodiments, oral azacitidine is not administered on Days 8through 14 (except for subjects in DL-2 who self-administer oralazacitidine at their assigned DL). Oral azacitidine followed by ABX 100mg/m² i.v. is administered on Day 15 in the clinic. On Days 16 through21, subjects self-administer oral azacitidine at their assigned DL.Subjects may continue to receive their assigned combination treatment ifthey have no unacceptable toxicity and if there is no clinical orradiographic evidence of disease progression. If combination treatmentis suspended for unacceptable toxicity that is believed to be related toABX, subjects may continue to take single agent oral azacitidine attheir assigned DL once the toxicity resolves.

Part 1/Arm C:

Subjects in Arm C receive their first dose of oral azacitidine at theirassigned DL in the clinic on Cycle 1 Day 1. Predose tumor collection(optional) accompanies oral azacitidine on Cycle 1 Days 1 and 15.Pre-dose PD blood collection (mandatory) accompanies oral azacitidinedosing on Cycle 1 Days 1, 8, and 15 and Cycle 2 Day 1. On Days 2 through7, 9 through 14 and 16 through 21 of each Cycle, subjectsself-administer oral azacitidine at their assigned DL; subjects in DL-1and DL-2 only self-administer oral azacitidine Days 2 through 7 and 9through 14 (FIG. 3).

In Cycle 2 and beyond, Arm C subjects self-administer oral azacitidineon Days 2 through 21; subjects in DL-1 and DL-2 only self-administeroral azacitidine on Days 2 through 14. Subjects who complete Cycle 2without unacceptable toxicity and without objective evidence of diseaseprogression on a tumor assessment may proceed to Cycle 3. Subjects maycontinue to receive oral azacitidine at their assigned DL as long asthey have no unacceptable toxicity and no clinical or radiographicevidence of disease progression.

DLT Definitions:

Any non-hematologic toxicity of NCI CTCAE v 4.0 Grade=3 that is believedto be related to oral azacitidine or to the combination of oralazacitidine with CBDCA or ABX with the following exceptions: (1) Grade 3emesis that responds to optimal antiemetic therapy within 72 hours; (2)Grade 3 diarrhea that responds to optimal medical management within 72hours; (3) Alopecia of any grade; (4) Grade 3 fatigue in a subject whohad Grade 2 fatigue at study entry and that recovers to baseline gradeor less within 72 hours; and (5) Grade 3 or 4 laboratory abnormalitiesthat are not accompanied by clinical signs or symptoms and are notbelieved by the investigator to be medically significant.

The following hematologic toxicities are considered DLT: (1) Grade 4neutropenia lasting>7 days or accompanied by fever; (2) Grade 3thrombocytopenia with clinically significant bleeding; and (3) Failureto meet hematologic criteria for starting Cycle 2 within 7 days of Cycle1 Day 28.

Definition of DLT-Evaluable Subjects:

To be evaluable for DLT for the purpose of dose escalation decisions, asubject must meet one of the following conditions: (1) Experienced a DLTduring Cycle 1; or (2) Did not receive Cycle 2 Day 1 treatment duesolely to not meeting hematologic criteria within 7 days of Cycle 1 Day28 (for Arm C subjects, Cycle 1 Day 21); or (3) Completed dosing forCycle 1 Day 28 (for Arm C subjects, Cycle 1 Day 21) without DLT and (i)missed no more than 4 total planned doses of oral azacitidine withinCycle 1; (ii) Arm A subjects: received scheduled dose of CBDCA duringCycle 1; and (iii) Arm B subjects: received all scheduled doses of ABXduring Cycle 1.

Subjects who do not meet any of the criteria for being DLT evaluable(e.g., who withdraw from study prior to the end of Cycle 1 for reasonsother than DLT) are replaced so that dose escalation decisions can bebased on a minimum of 6 DLT-evaluable subjects.

Part 2:

Once the RP2D and schedule have been determined for oral azacitidine asa single agent and in combination with CBDCA and/or ABX in Part 1,enrollment of Part 2 of the study begins. One objective of Part 2 is tofurther define the safety, PK, and PD of oral azacitidine combinationswith CBDCA and/or ABX and as a single agent in subjects with particulartumor types and to explore candidate predictive biomarkers of anti-tumoractivity. Up to 2 tumor types are examined for each Arm of the study.The definitive selection of tumor types evaluated in Part 2 aredetermined by any antitumor signal observed in Part 1. For each tumortype, enrollment proceeds in a 2-stage fashion. For each Arm, if atleast 2 objective responses are seen by Cycle 6 in the first 14subjects, an additional 6 subjects are enrolled for a total of 20subjects. If none of the first 14 subjects has an objective response, nofurther subject is enrolled.

PD and Predictive Biomarkers:

One objective of this study is to identify a dose and schedule of oralazacitidine that is not only safe but that exhibits pharmacologicactivity. Methylation changes in nucleated blood cells can provideconfirmation that a dose is pharmacologically active and can helpdistinguish which dose and schedule shows the most compellingpharmacologic activity.

Predictive biomarkers can allow prospective identification of patientswho are likely to benefit clinically from the combination of oralazacitidine as a single agent or combined with CBDCA or ABX. The PD andpredictive biomarkers analyzed in this study (e.g., Part 1 or Part 2 ofthe study) are shown in Table 4.

TABLE 4 PD and Predictive Biomarker Studies Tissue Analyte Assay PD^(a)Pred^(b) Whole Blood Genomic DNA Global Methylation X (PBMC) AnalysisRNA Global Methylation X Analysis Plasma Free DNA Candidate Gene X XMethylation Analysis Tumor (Fresh Genomic DNA Global Methylation X XFrozen) Analysis Candidate Gene X X Methylation Analysis RNA GlobalMethylation X X Analysis Candidate Gene Methylation Analysis Tumor(FFPE) Protein (IHC) DNMT1 X X Candidate Short X X Half-Life Proteins,DNA Damage, Apoptosis Markers ^(a)Change from pre-treatment to Cycle 1Day 7 ^(b)Baseline profile and change from pre-treatment to Cycle 1 Day7

Dosage Forms and Study Treatments:

Oral azacitidine is provided as 100 mg tablets for oral administration,for example, supplied by Celgene Corporation. See, e.g., U.S. PatentPublication No. 2009/0286752 (application Ser. No. 12/466,213), which isincorporated herein in its entirety.

Abraxane® is provided in single-use vials, for example, supplied byCelgene Corporation. Each single-use 50 mL vial contains 100 mgpaclitaxel and human albumin (HA) as a stabilizer. Unreconstituted ABXis stored at controlled room temperature (25° C. or 77° F.; excursionspermitted to 15-30° C.). Reconstituted ABX is refrigerated at 2° C. to8° C. (36° F. to 46° F.) and used within 8 hours. Both forms are storedin an area free of environmental extremes.

CBCDA may be obtained as a commercially available product through ahospital pharmacy or licensed distributor.

Each dose of oral azacitidine is taken with 8 ounces (240 mL) of roomtemperature water. Oral azacitidine may be taken on an empty stomach orwith food. If the dose is taken in the morning, subjects may consumetheir usual breakfast before or after administration.

No adjustment of the oral azacitidine dose is allowed during Cycle 1.Oral azacitidine may be held for up to 7 days between the end of Cycle 1and the start of Cycle 2 (to allow hematologic criteria) for Cycle 2 tobegin. For subjects who experience unacceptable toxicity after the startof Cycle 2, oral azacitidine may be held for up to 7 days or until thetoxicity recovers to grade 1 or less. If recovery has not occurred after7 days, dosing is permanently discontinued. Subjects who recover withinthe 7 day period may resume dosing at a reduced dose on the plannedCycle day (i.e., missed doses are not made up). For the first episode ofunacceptable toxicity in Cycle 2 or a later Cycle, if the subjectrecovers within 7 days of cessation of dosing with oral azacitidine andhad previously been receiving 300 mg of oral azacitidine, the subjectmay resume dosing at a dose of 200 mg. If the subject had previouslybeen receiving 200 mg of oral azacitidine, the subject may resume at adose of 100 mg. Subjects who experience unacceptable toxicity afterCycle 2 at a dose of 100 mg may resume dosing at the same dose if theyrecover within 7 days of dosing cessation.

For the second episode of unacceptable toxicity after Cycle 2, if thesubject recovers within 7 days of cessation of dosing and had previouslybeen receiving 200 mg of oral azacitidine, the subject may resume dosingat a dose of 100 mg. For subjects on reduced doses of oral azacitidine,the dose may be re-escalated (one dose level at a time) to theiroriginally assigned DL provided they have not experienced unacceptabletoxicity in 2 consecutive Cycles.

If, prior to the second episode of unacceptable toxicity, the subjecthad been receiving 100 mg of oral azacitidine, dosing is permanentlydiscontinued. Any subject who experiences a third episode ofunacceptable toxicity on a reduced dose of oral azacitidine discontinuesdosing permanently. No intra-subject dose escalation beyond the doseoriginally prescribed is allowed.

For the purposes of dose adjustment, unacceptable toxicity is defined asany AE that is deemed by the investigator to be related to oralazacitidine and/or to the combination of oral azacitidine with CBDCA orABX and that poses a medical risk or substantial discomfort to thesubject including but not limited to Grade 3 or 4 hematologic ornon-hematologic toxicity. If the unacceptable toxicity is believed bythe investigator to be more likely to be associated with the backboneagent (e.g., neuropathy with ABX), the subject may continue on singleagent oral azacitidine.

Administration of Oral Azacitidine:

Subjects are advised not to consume any grapefruit/grapefruit juiceduring the study, beginning 3 days prior to Cycle 1 Day 1. Subjectsdrink 8 ounces (240 mL) of room temperature water with each dose. Oralazacitidine may be taken on an empty stomach or with food. If the doseis taken in the morning, subjects may consume their usual breakfastbefore or after administration. The breakfast meal is not to exceed 600calories; however, the actual calorie count need not be measured orrecorded. If a meal other than breakfast is consumed, a light meal (notmore than 25% of a subject's usual daily calories) may be eaten beforeor after dose administration.

On days when subjects are not in the clinic, subjects take oralazacitidine at home. Subjects are given sufficient quantity of oralazacitidine for the dosing days at home. Subjects are instructed toinspect each oral azacitidine tablet and only take tablets that aretotally intact. Subjects are instructed to return any tablet found tonot be intact. Subjects are instructed to record the date and time oforal azacitidine administration in a Diary Card. On days when oralazacitidine is taken at home or on days when PK samples are notcollected during the clinic visit, subjects are encouraged to ingestoral azacitidine on an empty stomach or with food, with 8 ounces (240mL) of room temperature water.

Study Results:

In one embodiment, in Part 1, Arm A of the study, 5-azacytidine wasdosed from Day 1 to Day 14 and CBDCA was dosed on Day 8 at AUC 4, in a21-day cycle. Safety and PD were analyzed. In one embodiment, certainpatients were dosed with 5-azacytidine at a dose of 200 mg from Day 1 toDay 14 and CBDCA at a dose of AUC 4 on Day 8, in a 21-day cycle, totreat cancers, such as, NSCLC (non-small cell lung cancer), sarcoma, CRC(colorectal cancer), melanoma, ovarian cancer, or cervical cancer. Inone embodiment, certain patients were dosed with 5-azacytidine at a doseof 300 mg from Day 1 to Day 14 and CBDCA at a dose of AUC 4 on Day 8, ina 21-day cycle, to treat cancers, such as, mesothelioma, endometrialcancer, merkel cell cancer, melanoma, chodrosarcoma, NSCLC, or HNSCC(head and neck squamous cell carcinoma).

In one embodiment, in Part 1, Arm B of the study, 5-azacytidine wasdosed from Day 1 to Day 14 and ABX was dosed on Days 8 and 15 at 100mg/m², in a 21-day cycle. Safety and PD were analyzed. In oneembodiment, certain patients were dosed with 5-azacytidine at a dose of200 mg from Day 1 to Day 14 and ABX at a dose of 100 mg/m² on Days 8 andweekly thereafter, in a 21-day cycle, to treat cancers, such as,endometrial cancer, pancreatic cancer, ovarian cancer, or breast cancer.Partial responses were observed in endometrial cancer and pancreaticcancer (e.g., metastatic pancreatic cancer). For example, in one patientwith metastatic pancreatic cancer, after Cycle 2, CA19-9 level wasdecreased from 1867 to 15, and partial response was observed for sevenmonths or more. One patient with endometrial cancer progressed 8 monthson the study after 5 cycles of Carbo/Taxol, with no evidence of diseaseat primary site. In one embodiment, certain patients were dosed with5-azacytidine at a dose of 200 mg from Day 1 to Day 14 and ABX at a doseof 100 mg/m² on Days 8 and 15, in a 21-day cycle, to treat cancers, suchas, pancreatic cancer, cholangio cancer, HNSCC, CRC, or ovarian cancer.In one embodiment, certain patients were dosed with 5-azacytidine at adose of 300 mg from Day 1 to Day 14 and ABX at a dose of 100 mg/m² onDays 8 and 15, in a 21-day cycle, to treat cancers, such as, cholangiocancer, pancreatic cancer, or cervical cancer. Partial responses wereobserved in cervical cancer.

In one embodiment, in Part 1, Arm C of the study, 5-azacytidine wasdosed from Day 1 to Day 21, in a 21-day cycle (continuous). Safety andPD were analyzed. In one embodiment, certain patients were dosed with5-azacytidine at a dose of 200 mg from Day 1 to Day 21, in a 21-daycycle, to treat cancers, such as, CRC, head and neck cancer, or GIST(gastrointestinal stromal tumor). In one embodiment, certain patientswere dosed with 5-azacytidine at a dose of 300 mg from Day 1 to Day 21,in a 21-day cycle, to treat cancers, such as, CRC, NSCLC, or NP(nasopharyngeal) cancer. Partial responses were observed innasopharyngeal cancer.

Additional clinical efficacy observed in Part 1 of the study aresummarized below:

Prior Time Regimens AZA on (best response, ARM Dose Tumor Response Studymonths on Rx) Arm A 200 NSCLC SD >7 Erlotinib + (AZA + mg mo bevacizumabCBDCA) (SD 16 mo) 200 Sarcoma SD >7 Dox/Ifos (SD 3 mo) mg moGem/Taxotere (PD 2 mo) 300 Endometrial CA-125 ↓ >4 Carbo/Taxol mg >100%mo (SD 5 mo) Arm B 200 Pancreatic Mixed >3 Gem/Reolysin (AZA + mgresponse mo (SD 8 mo) ABX) 200 Endometrial PR ~8 Carbo/Taxol mg mo (SD 3mo) CC-122 (PD 1 mo) 200 Pancreatic PR >7 Gem mg mo (SD 7 mo) 200Pancreatic CA 19-9 ↓ ~4 Gem (PD 3 mo) mg  >50% mo Tivantiniberlotinib(PD 1 mo) 200 Colorectal 28% ↓ >3 5FU/Leuc/bev mg target mo (2 mo)refractory to prior treatment 200 Ovarian CA 125 ↓ >2 — mg  >50% mo ArmC 300 Nasopharyngeal PR >3 5FU/cisplat (AZA) mg mo (PR 5 mo) Erbitux (PD3 mo)

In one embodiment, in Part 2, Arm A of the study, 5-azacytidine wasdosed orally, e.g., at 300 mg (on Days 1 to 14 of a 21-day cycle), andCBDCA was dosed, e.g., at AUC 4, to treat patients with solid tumor,such as relapsed and refractory bladder cancer (e.g., bladder carcinoma,or urothelial malignancies) or relapsed and refractory ovarian cancer(e.g., epithelial ovarian carcinoma). Tissue samples are analyzed toevaluate activity and efficacy.

In one embodiment, in Part 2, Arm B of the study, 5-azacytidine wasdosed orally, e.g., at 200 mg (on Days 1 to 14 of a 21-day cycle), incombination with ABX (e.g., at a dose of 100 mg/m²), to treat patientswith solid tumor, such as relapsed and refractory NSCLC (non-small celllung cancer) or relapsed and refractory pancreatic cancer. Tissuesamples are analyzed to evaluate activity and efficacy.

In one embodiment, in Part 2, Arm C of the study, 5-azacytidine wasdosed alone (e.g., orally at a dose of 200 mg or 300 mg on Days 1 to 14of a 21-day cycle) to treat patients with solid tumor, such as relapsedand refractory colorectal cancer. Tissue samples are analyzed toevaluate activity and efficacy.

In one embodiment, in Part 1, Arm B of a study treating cancer patientswith 5-azacytidine and ABX, partial responses were seen in patients withpapillary serous endocervical adenocarcinoma, endometroid typewell-differentiated adenocarcinoma, metastatic ovarian carcinoma,pancreatic cancer, rectal adenocarcinoma, pancreatic carcinoma, ovariancancer, endometrial carcinoma, and metastatic pancreatic adenocarcinoma,as summarized below. Patients with pancreatic cancer, metastatic ovariancarcinoma, endometroid type well-differentiated adenocarcinoma, andpapillary serous endocervical adenocarcinoma showed a partial responsewith durations lasting between 4 and 8 months. See FIG. 7.

Best Percent Change in Tumor Cancer Size from Baseline (range) Papillaryserous endocervical −73 to −83 adenocarcinoma Endometroid type well- −64to −74 differentiated adenocarcinoma Metastatic ovarian carcinoma −37 to−47 Pancreatic cancer −34 to −44 Rectal adenocarcinoma −24 to −34Pancreatic carcinoma −24 to −34 Pancreatic carcinoma −21 to −31 Ovariancancer −18 to −28 Endometrial carcinoma  −8 to −18 Metastatic pancreatic −1 to −11 adenocarcinoma

In one embodiment, in Part 1, Arm C of a study treating cancer patientswith 5-azacytidine, partial responses were seen in patients withnasopharyngeal carcinoma, cervical adenocarcinoma, nonsmall celladenocarcinoma of the lung, colon adenocarcinoma, and colon Krasadenocarcinoma, as summarized below. See FIG. 8.

Best Percent Change in Tumor Size from Cancer Baseline (range)Nasopharyngeal carcinoma −29 to −39 Nasopharyngeal carcinoma −23 to −33Cervical adenocarcinoma −22 to −32 Nonsmall cell −13 to −23adenocarcinoma of the lung Colon adenocarcinoma  −9 to −19 Colon Krasadenocarcinoma  −1 to −11

B. Example 2

DNA methylation is employed as a biomarker to monitor responses inpatients treated with azacitidine in the clinical studies describedherein. Analysis is performed with an Infinium Assay (commerciallyavailable from Illumina, Inc., San Diego, Calif.). The Infinium Assaycombined with BeadChips allows large-scale interrogation of variationsin the human genome. For example, the Infinium HumanMethylation27BeadChip enables interrogation of 27,578 CpG loci, covering over 14,000genes. The DNA Methylation Assay protocol includes the following steps:(1) bisulfite conversion; (2) DNA amplification; (3) DNA fragmentation;(4) DNA precipitation; (5) DNA hybridization to BeadChip; (6) extensionand staining on BeadChip; and (7) imaging of BeadChip. In otherembodiments, DNA methylation assay with 450K array (instead of 27Karray) is used.

The assay for methylation is used to detect methylation status atindividual CpG loci by typing bisulfite-converted DNA. Methylationprotected C from conversion, whereas unmethylated C is converted to T. Apair of bead-bound probes is used to detect the presence of T or C byhybridization followed by single-base extension with a labelednucleotide. Up to twelve samples are profiled in parallel. Blood sampleswere collected and DNA methylation was analyzed in parallel. In otherembodiments, bone marrow samples are collected and DNA methylationanalyzed in parallel.

Methylation of plasma DNA and PBMC DNA of patients from Part I of theclinical study exemplified in Example 1 was analyzed.

C. Example 3

Clinical studies are conducted to assess the ability of an oralformulation comprising a cytidine analog, such as 5-azacytidine, totreat patients having lung cancer, e.g., non-small-cell lung cancer(NSCLC). Such studies may include, e.g., an assessment of the ability tostop or reverse the growth of particular NSCLC cell types in patientshaving NSCLC). In certain clinical studies, patients are tested forparticular NSCLC cell types, prior to administration of the oralformulation. In certain clinical studies, patients with cell types knownor believed to benefit preferentially from cytidine analog (e.g.,5-azacytidine) administration may be enrolled. In certain clinicalstudies, patients having NSCLC are enrolled without analysis ofparticular NSCLC cell type. In certain clinical studies, patients havingany type of NSCLC cells are candidates for treatment with an oralformulation provided herein.

In certain clinical studies, patients from any of the three main NSCLCgroups may be enrolled, i.e., (1) patients with tumors that aresurgically resectable; (2) patients with either locally or regionallyadvanced lung cancer; or (3) patients with distant metastases at thetime of diagnosis. In certain clinical studies, patients may becurrently undergoing additional treatment for NSCLC, including, e.g.,surgery, chemotherapy, or radiation therapy.

In certain clinical studies, patients who are administered an oralformulation comprising a cytidine analog (e.g., 5-azacytidine) may alsobe administered one or more additional therapeutic agents, examples ofwhich are disclosed herein. The additional therapeutic agent(s) may beadministered in the same oral formulation as the cytidine analog, or maybe co-administered (e.g., via PO, SC or IV administration) incombination with an oral formulation comprising the cytidine analog. Theappropriate amount and dosing schedule for an additional therapeuticagent may be determined for a particular patient using methods known inthe art.

In particular embodiments, the co-administered agent is carboplatin. Inparticular embodiments, the co-administered agent is paclitaxel (e.g.,Abraxane®).

An association between gene methylation and recurrence of NSCLC tumorsis known in the art. See, e.g., M. V. Brock et al., N. Engl. J. Med.,2008, 358(11):1118-28. Accordingly, in certain clinical studies providedherein, patients are screened prior to enrollment and/or monitoredduring the trial for DNA or RNA methylation levels, which indicate apotential response to treatment with an oral formulation comprising acytidine analog (e.g., 5-azacytidine). In certain clinical studies,patients with high levels of DNA methylation (e.g., CpG islandmethylation) and/or an increased potential for transcriptional silencingof tumor-suppressor genes may be administered a cytidine analog (e.g.,5-azacytidine) known or believed to prevent or reverse hypermethylation(e.g., by reducing the activity of one or more DNA methyltransferaseenzymes). In certain clinical studies, patients with high levels of DNAmethylation (e.g., CpG island methylation) of certain genes may beadministered a cytidine analog (e.g., 5-azacytidine). In certainclinical studies, patients with low levels of DNA methylation (e.g., CpGisland methylation) of certain genes may be administered a cytidineanalog (e.g., 5-azacytidine). In certain clinical studies, patients witha particular DNA methylation signature (e.g., CpG island methylation) ofcertain genes may be administered a cytidine analog (e.g.,5-azacytidine). In such studies, patients may also be co-administeredone or more additional therapeutic agents known or believed to reduceepigenetic silencing, such as, e.g., compounds that inhibit histonedeacetylase enzymes (HDACs), which regulate the acetylation anddeacetylation of histone residues that increase or decrease geneexpression. See, e.g., J. G. Herman & S. B. Baylin, N. Engl. J. Med.,2003, 349:2042-54; P. A. Jones & S. B. Baylin, Nature Rev. Gen., 2002,3:415-28. Suitable HDAC inhibitors for co-administration in the clinicalstudies disclosed herein are known in the art and/or described herein(e.g., entinostat or vorinostat).

The amount of cytidine analog (e.g., 5-azacytidine) in the oralformulations administered during the clinical studies depends, e.g., onthe individual characteristics of the patient, including, inter alia,the stage and progression of the patient's NSCLC, the patient's age andweight, the patient's prior treatment regimens, and other variables, asknown in the art. In certain clinical studies, potential starting dosesmay be, e.g., about 50 mg, about 60 mg, about 70 mg, about 75 mg, about80 mg, about 90 mg, about 100 mg, about 120 mg, about 140 mg, about 150mg, about 160 mg, about 180 mg, about 200 mg, about 220 mg, about 240mg, about 250 mg, about 300 mg, about 350 mg, about 360 mg, about 400mg, about 420 mg, about 450 mg, about 480 mg, about 500 mg, about 540mg, about 600 mg, about 660 mg, about 720 mg, about 780 mg, about 840mg, about 900 mg, about 960 mg, about 1020 mg, or greater than about1020 mg of the cytidine analog (e.g., 5-azacytidine) daily for aspecified time period, e.g., about 1 week, about 1.5 weeks, about 2weeks, about 2.5 weeks, about 3 weeks, about 3.5 weeks, about 1 month,about 1.5 months, about 2 months, or a longer time period. Otherpotential starting doses and time periods are disclosed herein. Cyclesmay be repeated as desired, e.g., over a period of one or more months,as disclosed herein. After a certain number of cycles, the dosage may beincreased to increase the beneficial effect, provided such an increasedoes not cause undesirable toxicity effects. Patients may be treated fora minimum number of cycles, as disclosed herein. Complete or partialresponse may require additional treatment cycles. Treatment may becontinued as long as the patient continues to benefit.

D. Example 4

Clinical studies are conducted to assess the ability of an oralformulation comprising a cytidine analog, such as 5-azacytidine, totreat patients having an ovarian cancer (including, e.g., the ability tostop or reverse the growth of cancer cells in patients having an ovariancancer). Particular ovarian cancers include, but are not limited to,ovarian epithelial cancer, ovarian germ cell tumors, and ovarian lowmalignant potential tumors. In certain clinical studies, patients arescreened for the presence of a particular type of ovarian cancer priorto administration of the oral formulation. In certain clinical studies,patients with a type of ovarian cancer known or believed to benefitpreferentially from cytidine analog (e.g., 5-azacytidine) administrationmay be enrolled. In certain clinical studies, patients having ovariancancer are enrolled without screening for particular ovarian cancertypes. In certain clinical studies, patients having any type of ovariancancer are candidates for treatment with an oral formulation providedherein. In certain clinical studies, patients may be currentlyundergoing additional treatment for ovarian cancer, including, e.g.,surgery, chemotherapy, or radiation therapy.

In certain clinical studies, patients who are administered an oralformulation comprising a cytidine analog (e.g., 5-azacytidine) may alsobe administered one or more additional therapeutic agents, examples ofwhich are disclosed herein (e.g., carboplatin). The additionaltherapeutic agent(s) may be administered in the same oral formulation asthe cytidine analog, or may be co-administered (e.g., via PO, SC or IVadministration) in combination with an oral formulation comprising acytidine analog. The appropriate amount and dosing schedule for anadditional therapeutic agent may be determined for a particular patientusing methods known in the art.

In particular embodiments, the co-administered agent is carboplatin. Inparticular embodiments, the co-administered agent is paclitaxel (e.g.,Abraxane®).

An association between gene methylation and ovarian cancer is known inthe art. See, e.g., G. Gifford et al., Clin. Cancer Res., 2004,10:4420-26. Accordingly, in certain clinical studies provided herein,patients are screened prior to enrollment and/or monitored during thetrial for DNA or RNA methylation levels, which indicate a potentialresponse to treatment with an oral formulation comprising a cytidineanalog (e.g., 5-azacytidine). In certain clinical studies, patients withhigh levels of DNA methylation (e.g., CpG island methylation) and/or anincreased potential for transcriptional silencing of tumor-suppressorgenes may be administered a cytidine analog (e.g., 5-azacytidine) knownor believed to prevent or reverse hypermethylation (e.g., by reducingthe activity of one or more DNA methyltransferase enzymes). In certainclinical studies, patients with high levels of DNA methylation (e.g.,CpG island methylation) of certain genes may be administered a cytidineanalog (e.g., 5-azacytidine). In certain clinical studies, patients withlow levels of DNA methylation (e.g., CpG island methylation) of certaingenes may be administered a cytidine analog (e.g., 5-azacytidine). Incertain clinical studies, patients with a particular DNA methylationsignature (e.g., CpG island methylation) of certain genes may beadministered a cytidine analog (e.g., 5-azacytidine). In such studies,patients may also be co-administered one or more additional therapeuticagents known or believed to reduce epigenetic silencing, such as, e.g.,compounds that inhibit histone deacetylase enzymes (HDACs), whichregulate the acetylation and deacetylation of histone residues thatincrease or decrease gene expression. See, e.g., J. G. Herman & S. B.Baylin, N. Engl. J. Med., 2003, 349:2042-54; P. A. Jones & S. B. Baylin,Nature Rev. Gen., 2002, 3:415-28. Suitable HDAC inhibitors forco-administration in the clinical studies disclosed herein are known inthe art and/or described herein (e.g., entinostat or vorinostat).

The amount of cytidine analog (e.g., 5-azacytidine) in the oralformulations administered during the clinical studies depends, e.g., onthe individual characteristics of the patient, including, inter alia,the type, stage, and progression of the patient's ovarian cancer, thepatient's age and weight, the patient's prior treatment regimens, andother variables, as known in the art. n certain clinical studies,potential starting doses may be, e.g., about 50 mg, about 60 mg, about70 mg, about 75 mg, about 80 mg, about 90 mg, about 100 mg, about 120mg, about 140 mg, about 150 mg, about 160 mg, about 180 mg, about 200mg, about 220 mg, about 240 mg, about 250 mg, about 300 mg, about 350mg, about 360 mg, about 400 mg, about 420 mg, about 450 mg, about 480mg, about 500 mg, about 540 mg, about 600 mg, about 660 mg, about 720mg, about 780 mg, about 840 mg, about 900 mg, about 960 mg, about 1020mg, or greater than about 1020 mg of the cytidine analog (e.g.,5-azacytidine) daily for a specified time period, e.g., about 1 week,about 1.5 weeks, about 2 weeks, about 2.5 weeks, about 3 weeks, about3.5 weeks, about 1 month, about 1.5 months, about 2 months, or a longertime period. Other potential starting doses and time periods aredisclosed herein. Cycles may be repeated as desired, e.g., over a periodof one or more months, as disclosed herein. After a certain number ofcycles, the dosage may be increased to increase the beneficial effect,provided such an increase does not cause undesirable toxicity effects.Patients may be treated for a minimum number of cycles, as disclosedherein. Complete or partial response may require additional treatmentcycles. Treatment may be continued as long as the patient continues tobenefit.

E. Example 5

Clinical studies are conducted to assess the ability of an oralformulation comprising a cytidine analog, such as 5-azacytidine, totreat patients having a pancreatic cancer (including, e.g., the abilityto stop or reverse the growth of cancer cells in patients havingpancreatic cancer). In certain clinical studies, patients are screenedprior to enrollment for a particular type of pancreatic cancer prior toadministration of the oral formulation. Cellular classifications ofpancreatic cancers are known in the art and include, e.g., duct cellcarcinoma; acinar cell carcinoma; papillary mucinous carcinoma; signetring carcinoma; adenosquamous carcinoma; undifferentiated carcinoma;mucinous carcinoma; giant cell carcinoma; mixed type (ductal-endocrineor acinar-endocrine); small cell carcinoma; cystadenocarcinoma (serousand mucinous types); unclassified; pancreatoblastoma; papillary-cysticneoplasm (Frantz tumor); invasive adenocarcinoma associated with cysticmucinous neoplasm or intraductal papillary mucinous neoplasm; mucinouscystic tumor with dysplasia; intraductal papillary mucinous tumor withdysplasia; and pseudopapillary solid tumor. In certain clinical studies,patients are screened prior to enrollment for a particular stage ofpancreatic cancer (e.g., the size of the tumor in the pancreas, whetherthe cancer has spread, and if so, to what parts of the body) prior toadministration of the oral formulation. In certain clinical studies,pancreatic cancer patients believed to benefit preferentially fromcytidine analog (e.g., 5-azacytidine) administration may be enrolled. Incertain clinical studies, patients having pancreatic cancer are enrolledwithout screening for particular pancreatic cancer types. In certainclinical studies, patients having any type of pancreatic cancer arecandidates for treatment with an oral formulation provided herein. Incertain clinical studies, patients may be currently undergoingadditional treatment for pancreatic cancer, including, e.g., surgery,chemotherapy, or radiation therapy.

In certain clinical studies, patients who are administered an oralformulation comprising a cytidine analog (e.g., 5-azacytidine) may alsobe administered one or more additional therapeutic agents, examples ofwhich are disclosed herein (e.g., gemcitabine). The additionaltherapeutic agent(s) may be administered in the same oral formulation asthe cytidine analog, or may be co-administered (e.g., via PO, SC or IVadministration) in combination with an oral formulation comprising acytidine analog. The appropriate amount and dosing schedule for anadditional therapeutic agent may be determined for a particular patientusing methods known in the art.

In particular embodiments, the co-administered agent is carboplatin. Inparticular embodiments, the co-administered agent is paclitaxel (e.g.,Abraxane®).

In certain clinical studies provided herein, patients are screened priorto enrollment and/or monitored during the trial for DNA or RNAmethylation levels, which indicate a potential response to treatmentwith an oral formulation comprising a cytidine analog (e.g.,5-azacytidine). In certain clinical studies, patients with high levelsof DNA methylation (e.g., CpG island methylation) and/or an increasedpotential for transcriptional silencing of tumor-suppressor genes may beadministered a cytidine analog (e.g., 5-azacytidine) known or believedto prevent or reverse hypermethylation (e.g., by reducing the activityof one or more DNA methyltransferase enzymes). In certain clinicalstudies, patients with high levels of DNA methylation (e.g., CpG islandmethylation) of certain genes may be administered a cytidine analog(e.g., 5-azacytidine). In certain clinical studies, patients with lowlevels of DNA methylation (e.g., CpG island methylation) of certaingenes may be administered a cytidine analog (e.g., 5-azacytidine). Incertain clinical studies, patients with a particular DNA methylationsignature (e.g., CpG island methylation) of certain genes may beadministered a cytidine analog (e.g., 5-azacytidine). In such studies,patients may also be co-administered one or more additional therapeuticagents known or believed to reduce epigenetic silencing, such as, e.g.,compounds that inhibit histone deacetylase enzymes (HDACs), whichregulate the acetylation and deacetylation of histone residues thatincrease or decrease gene expression. See, e.g., J. G. Herman & S. B.Baylin, N. Engl. J. Med., 2003, 349:2042-54; P. A. Jones & S. B. Baylin,Nature Rev. Gen., 2002, 3:415-28. Suitable HDAC inhibitors forco-administration in the clinical studies disclosed herein are known inthe art and/or described herein (e.g., entinostat or vorinostat).

The amount of cytidine analog (e.g., 5-azacytidine) in the oralformulations administered during the clinical studies depends, e.g., onthe individual characteristics of the patient, including, inter alia,the type, stage, and progression of the patient's pancreatic cancer, thepatient's age and weight, the patient's prior treatment regimens, andother variables, as known in the art. In certain clinical studies,potential starting doses may be, e.g., about 50 mg, about 60 mg, about70 mg, about 75 mg, about 80 mg, about 90 mg, about 100 mg, about 120mg, about 140 mg, about 150 mg, about 160 mg, about 180 mg, about 200mg, about 220 mg, about 240 mg, about 250 mg, about 300 mg, about 350mg, about 360 mg, about 400 mg, about 420 mg, about 450 mg, about 480mg, about 500 mg, about 540 mg, about 600 mg, about 660 mg, about 720mg, about 780 mg, about 840 mg, about 900 mg, about 960 mg, about 1020mg, or greater than about 1020 mg of the cytidine analog (e.g.,5-azacytidine) daily for a specified time period, e.g., about 1 week,about 1.5 weeks, about 2 weeks, about 2.5 weeks, about 3 weeks, about3.5 weeks, about 1 month, about 1.5 months, about 2 months, or a longertime period. Other potential starting doses and time periods aredisclosed herein. Cycles may be repeated as desired, e.g., over a periodof one or more months, as disclosed herein. After a certain number ofcycles, the dosage may be increased to increase the beneficial effect,provided such an increase does not cause undesirable toxicity effects.Patients may be treated for a minimum number of cycles, as disclosedherein. Complete or partial response may require additional treatmentcycles. Treatment may be continued as long as the patient continues tobenefit.

F. Example 6

Clinical studies are conducted to assess the ability of an oralformulation comprising a cytidine analog, such as 5-azacytidine, totreat patients having a colorectal cancer (including, e.g., the abilityto stop or reverse the growth of cancer cells in patients having acolorectal cancer). In certain clinical studies, patients are screenedprior to enrollment for a particular type of colorectal cancer prior toadministration of the oral formulation. Histologic types of coloncancers are known in the art and include, e.g., adenocarcinoma; mucinous(colloid) adenocarcinoma; signet ring adenocarcinoma; scirrhous tumors;and neuroendocrine tumors. The World Health Organization classificationof tumors of the colon and rectum include (1) Epithelial Tumors, whichinclude: Adenoma (e.g., tubular, villous, tubulovillous, and serrated);Intraepithelial neoplasia (dysplasia) associated with chronicinflammatory diseases (e.g., low-grade glandular intraepithelialneoplasia and high-grade glandular intraepithelial neoplasia); Carcinoma(e.g., adenocarcinoma, mucinous adenocarcinoma, signet-ring cellcarcinoma, small cell carcinoma, adenosquamous carcinoma, medullarycarcinoma, and undifferentiated carcinoma); Carcinoid(well-differentiated neuroendocrine neoplasm) (e.g., enterochromaffin(EC)-cell, serotonin-producing neoplasm, L-cell, glucagon-like peptideand pancreatic polypeptide/peptide YY (PYY)-producing tumor, andothers); and Mixed carcinoma-adenocarcinoma; and (2) NonepithelialTumors, which include: Lipoma; Leiomyoma; Gastrointestinal stromaltumor; Leiomyosarcoma; Angiosarcoma; Kaposi sarcoma; Melanoma; andothers; as well as Malignant lymphomas (e.g., marginal zone B-celllymphoma of mucosa-associated lymphoid tissue type, mantle celllymphoma, diffuse large B-cell lymphoma, Burkitt lymphoma, andBurkitt-like/atypical Burkitt lymphoma. In certain clinical studies,patients are screened prior to enrollment for a particular stage ofcolorectal cancer (e.g., the size of the tumor in the colon or rectum,whether the cancer has spread, and if so, to what parts of the body)prior to administration of the oral formulation. In certain clinicalstudies, colorectal cancer patients believed to benefit preferentiallyfrom cytidine analog (e.g., 5-azacytidine) administration may beenrolled. In certain clinical studies, patients having a colorectalcancer are enrolled without screening for particular colorectal cancertypes. In certain clinical studies, patients having any type ofcolorectal cancer are candidates for treatment with an oral formulationprovided herein. In certain clinical studies, patients may be currentlyundergoing additional treatment for colorectal cancer, including, e.g.,surgery, chemotherapy, or radiation therapy.

In certain clinical studies, patients who are administered an oralformulation comprising a cytidine analog (e.g., 5-azacytidine) may alsobe administered one or more additional therapeutic agents, examples ofwhich are disclosed herein. The additional therapeutic agent(s) may beadministered in the same oral formulation as the cytidine analog, or maybe co-administered (e.g., via PO, SC or IV administration) incombination with an oral formulation comprising a cytidine analog. Theappropriate amount and dosing schedule for an additional therapeuticagent may be determined for a particular patient using methods known inthe art.

In particular embodiments, the co-administered agent is carboplatin. Inparticular embodiments, the co-administered agent is paclitaxel (e.g.,Abraxane®).

An association between gene methylation and colorectal cancer is knownin the art. See, e.g., A. M. Jubb et al., J. Pathol., 2001, 195:111-134.Accordingly, in certain clinical studies provided herein, patients arescreened prior to enrollment and/or monitored during the trial for DNAor RNA methylation levels, which indicate a potential response totreatment with an oral formulation comprising a cytidine analog (e.g.,5-azacytidine). In certain clinical studies, patients with high levelsof DNA methylation (e.g., CpG island methylation) and/or an increasedpotential for transcriptional silencing of tumor-suppressor genes may beadministered a cytidine analog (e.g., 5-azacytidine) known or believedto prevent or reverse hypermethylation (e.g., by reducing the activityof one or more DNA methyltransferase enzymes). In certain clinicalstudies, patients with high levels of DNA methylation (e.g., CpG islandmethylation) of certain genes may be administered a cytidine analog(e.g., 5-azacytidine). In certain clinical studies, patients with lowlevels of DNA methylation (e.g., CpG island methylation) of certaingenes may be administered a cytidine analog (e.g., 5-azacytidine). Incertain clinical studies, patients with a particular DNA methylationsignature (e.g., CpG island methylation) of certain genes may beadministered a cytidine analog (e.g., 5-azacytidine). In such studies,patients may also be co-administered one or more additional therapeuticagents known or believed to reduce epigenetic silencing, such as, e.g.,compounds that inhibit histone deacetylase enzymes (HDACs), whichregulate the acetylation and deacetylation of histone residues thatincrease or decrease gene expression. See, e.g., J. G. Herman & S. B.Baylin, N. Engl. J. Med., 2003, 349:2042-54; P. A. Jones & S. B. Baylin,Nature Rev. Gen., 2002, 3:415-28. Suitable HDAC inhibitors forco-administration in the clinical studies disclosed herein are known inthe art and/or described herein (e.g., entinostat or vorinostat).

The amount of cytidine analog (e.g., 5-azacytidine) in the oralformulations administered during the clinical studies depends, e.g., onthe individual characteristics of the patient, including, inter alia,the type, stage, and progression of the patient's colorectal cancer, thepatient's age and weight, the patient's prior treatment regimens, andother variables, as known in the art. In certain clinical studies,potential starting doses may be, e.g., about 50 mg, about 60 mg, about70 mg, about 75 mg, about 80 mg, about 90 mg, about 100 mg, about 120mg, about 140 mg, about 150 mg, about 160 mg, about 180 mg, about 200mg, about 220 mg, about 240 mg, about 250 mg, about 300 mg, about 350mg, about 360 mg, about 400 mg, about 420 mg, about 450 mg, about 480mg, about 500 mg, about 540 mg, about 600 mg, about 660 mg, about 720mg, about 780 mg, about 840 mg, about 900 mg, about 960 mg, about 1020mg, or greater than about 1020 mg of the cytidine analog (e.g.,5-azacytidine) daily for a specified time period, e.g., about 1 week,about 1.5 weeks, about 2 weeks, about 2.5 weeks, about 3 weeks, about3.5 weeks, about 1 month, about 1.5 months, about 2 months, or a longertime period. Other potential starting doses and time periods aredisclosed herein. After a certain number of cycles, the dosage may beincreased to increase the beneficial effect, provided such an increasedoes not cause undesirable toxicity effects. Patients may be treated fora minimum number of cycles, as disclosed herein. Complete or partialresponse may require additional treatment cycles. Treatment may becontinued as long as the patient continues to benefit.

G. Example 7

Clinical studies are conducted to assess the ability of an oralformulation comprising a cytidine analog, such as 5-azacytidine, totreat patients having a bladder cancer (including, e.g., the ability tostop or reverse the growth of cancer cells in patients having a bladdercancer). In certain clinical studies, patients are screened prior toenrollment for a particular type of bladder cancer prior toadministration of the oral formulation. In certain clinical studies,patients are screened prior to enrollment for a particular stage ofbladder cancer (e.g., the size of the tumor, whether the cancer hasspread, and if so, to what parts of the body) prior to administration ofthe oral formulation. In certain clinical studies, patients are screenedprior to enrollment for a particular type of bladder cancer prior toadministration of the oral formulation. In certain clinical studies,bladder cancer patients believed to benefit preferentially from cytidineanalog (e.g., 5-azacytidine) administration may be enrolled. In certainclinical studies, patients having a bladder cancer are enrolled withoutscreening for particular bladder cancer types. In certain clinicalstudies, patients having any type of bladder cancer are candidates fortreatment with an oral formulation provided herein. In certain clinicalstudies, patients may be currently undergoing additional treatment forbladder cancer, including, e.g., surgery, chemotherapy, or radiationtherapy.

In certain clinical studies, patients who are administered an oralformulation comprising a cytidine analog (e.g., 5-azacytidine) may alsobe administered one or more additional therapeutic agents, examples ofwhich are disclosed herein. The additional therapeutic agent(s) may beadministered in the same oral formulation as the cytidine analog, or maybe co-administered (e.g., via PO, SC or IV administration) incombination with an oral formulation comprising a cytidine analog. Theappropriate amount and dosing schedule for an additional therapeuticagent may be determined for a particular patient using methods known inthe art.

In particular embodiments, the co-administered agent is carboplatin. Inparticular embodiments, the co-administered agent is paclitaxel (e.g.,Abraxane®).

In certain clinical studies provided herein, patients are screened priorto enrollment and/or monitored during the trial for DNA or RNAmethylation levels, which indicate a potential response to treatmentwith an oral formulation comprising a cytidine analog (e.g.,5-azacytidine). In certain clinical studies, patients with high levelsof DNA methylation (e.g., CpG island methylation) and/or an increasedpotential for transcriptional silencing of tumor-suppressor genes may beadministered a cytidine analog (e.g., 5-azacytidine) known or believedto prevent or reverse hypermethylation (e.g., by reducing the activityof one or more DNA methyltransferase enzymes). In certain clinicalstudies, patients with high levels of DNA methylation (e.g., CpG islandmethylation) of certain genes may be administered a cytidine analog(e.g., 5-azacytidine). In certain clinical studies, patients with lowlevels of DNA methylation (e.g., CpG island methylation) of certaingenes may be administered a cytidine analog (e.g., 5-azacytidine). Incertain clinical studies, patients with a particular DNA methylationsignature (e.g., CpG island methylation) of certain genes may beadministered a cytidine analog (e.g., 5-azacytidine). In such studies,patients may also be co-administered one or more additional therapeuticagents known or believed to reduce epigenetic silencing, such as, e.g.,compounds that inhibit histone deacetylase enzymes (HDACs), whichregulate the acetylation and deacetylation of histone residues thatincrease or decrease gene expression. See, e.g., J. G. Herman & S. B.Baylin, N. Engl. J. Med., 2003, 349:2042-54; P. A. Jones & S. B. Baylin,Nature Rev. Gen., 2002, 3:415-28. Suitable HDAC inhibitors forco-administration in the clinical studies disclosed herein are known inthe art and/or described herein (e.g., entinostat or vorinostat).

The amount of cytidine analog (e.g., 5-azacytidine) in the oralformulations administered during the clinical studies depends, e.g., onthe individual characteristics of the patient, including, inter alia,the type, stage, and progression of the patient's bladder cancer, thepatient's age and weight, the patient's prior treatment regimens, andother variables, as known in the art. In certain clinical studies,potential starting doses may be, e.g., about 50 mg, about 60 mg, about70 mg, about 75 mg, about 80 mg, about 90 mg, about 100 mg, about 120mg, about 140 mg, about 150 mg, about 160 mg, about 180 mg, about 200mg, about 220 mg, about 240 mg, about 250 mg, about 300 mg, about 350mg, about 360 mg, about 400 mg, about 420 mg, about 450 mg, about 480mg, about 500 mg, about 540 mg, about 600 mg, about 660 mg, about 720mg, about 780 mg, about 840 mg, about 900 mg, about 960 mg, about 1020mg, or greater than about 1020 mg of the cytidine analog (e.g.,5-azacytidine) daily for a specified time period, e.g., about 1 week,about 1.5 weeks, about 2 weeks, about 2.5 weeks, about 3 weeks, about3.5 weeks, about 1 month, about 1.5 months, about 2 months, or a longertime period. Other potential starting doses and time periods aredisclosed herein. After a certain number of cycles, the dosage may beincreased to increase the beneficial effect, provided such an increasedoes not cause undesirable toxicity effects. Patients may be treated fora minimum number of cycles, as disclosed herein. Complete or partialresponse may require additional treatment cycles. Treatment may becontinued as long as the patient continues to benefit.

H. Example 8

Clinical studies are conducted to assess the ability of an oralformulation comprising a cytidine analog, such as 5-azacytidine, totreat patients having a breast cancer (including, e.g., the ability tostop or reverse the growth of cancer cells in patients having a breastcancer). In certain clinical studies, patients are screened prior toenrollment for a particular type of breast cancer prior toadministration of the oral formulation. In certain clinical studies,patients are screened prior to enrollment for a particular stage ofbreast cancer (e.g., the size of the tumor in the breast, whether thecancer has spread, and if so, to what parts of the body) prior toadministration of the oral formulation. In certain clinical studies,patients are screened prior to enrollment for a particular type ofbreast cancer prior to administration of the oral formulation. Incertain clinical studies, breast cancer patients believed to benefitpreferentially from cytidine analog (e.g., 5-azacytidine) administrationmay be enrolled. In certain clinical studies, patients having a breastcancer are enrolled without screening for particular breast cancertypes. In certain clinical studies, patients having any type of breastcancer are candidates for treatment with an oral formulation providedherein. In certain clinical studies, patients may be currentlyundergoing additional treatment for breast cancer, including, e.g.,surgery, chemotherapy, or radiation therapy.

In certain clinical studies, patients who are administered an oralformulation comprising a cytidine analog (e.g., 5-azacytidine) may alsobe administered one or more additional therapeutic agents, examples ofwhich are disclosed herein. The additional therapeutic agent(s) may beadministered in the same oral formulation as the cytidine analog, or maybe co-administered (e.g., via PO, SC or IV administration) incombination with an oral formulation comprising a cytidine analog. Theappropriate amount and dosing schedule for an additional therapeuticagent may be determined for a particular patient using methods known inthe art. In some embodiments, the co-administered agent is carboplatin.In particular embodiments, the co-administered agent is paclitaxel(e.g., Abraxane®).

In certain clinical studies provided herein, patients are screened priorto enrollment and/or monitored during the trial for DNA or RNAmethylation levels, which indicate a potential response to treatmentwith an oral formulation comprising a cytidine analog (e.g.,5-azacytidine). In certain clinical studies, patients with high levelsof DNA methylation (e.g., CpG island methylation) and/or an increasedpotential for transcriptional silencing of tumor-suppressor genes may beadministered a cytidine analog (e.g., 5-azacytidine) known or believedto prevent or reverse hypermethylation (e.g., by reducing the activityof one or more DNA methyltransferase enzymes). In certain clinicalstudies, patients with high levels of DNA methylation (e.g., CpG islandmethylation) of certain genes may be administered a cytidine analog(e.g., 5-azacytidine). In certain clinical studies, patients with lowlevels of DNA methylation (e.g., CpG island methylation) of certaingenes may be administered a cytidine analog (e.g., 5-azacytidine). Incertain clinical studies, patients with a particular DNA methylationsignature (e.g., CpG island methylation) of certain genes may beadministered a cytidine analog (e.g., 5-azacytidine). In such studies,patients may also be co-administered one or more additional therapeuticagents known or believed to reduce epigenetic silencing, such as, e.g.,compounds that inhibit histone deacetylase enzymes (HDACs), whichregulate the acetylation and deacetylation of histone residues thatincrease or decrease gene expression. See, e.g., J. G. Herman & S. B.Baylin, N. Engl. J. Med., 2003, 349:2042-54; P. A. Jones & S. B. Baylin,Nature Rev. Gen., 2002, 3:415-28. Suitable HDAC inhibitors forco-administration in the clinical studies disclosed herein are known inthe art and/or described herein (e.g., entinostat or vorinostat).

The amount of cytidine analog (e.g., 5-azacytidine) in the oralformulations administered during the clinical studies depends, e.g., onthe individual characteristics of the patient, including, inter alia,the type, stage, and progression of the patient's breast cancer, thepatient's age and weight, the patient's prior treatment regimens, andother variables, as known in the art. In certain clinical studies,potential starting doses may be, e.g., about 50 mg, about 60 mg, about70 mg, about 75 mg, about 80 mg, about 90 mg, about 100 mg, about 120mg, about 140 mg, about 150 mg, about 160 mg, about 180 mg, about 200mg, about 220 mg, about 240 mg, about 250 mg, about 300 mg, about 350mg, about 360 mg, about 400 mg, about 420 mg, about 450 mg, about 480mg, about 500 mg, about 540 mg, about 600 mg, about 660 mg, about 720mg, about 780 mg, about 840 mg, about 900 mg, about 960 mg, about 1020mg, or greater than about 1020 mg of the cytidine analog (e.g.,5-azacytidine) daily for a specified time period, e.g., about 1 week,about 1.5 weeks, about 2 weeks, about 2.5 weeks, about 3 weeks, about3.5 weeks, about 1 month, about 1.5 months, about 2 months, or a longertime period. Other potential starting doses and time periods aredisclosed herein. After a certain number of cycles, the dosage may beincreased to increase the beneficial effect, provided such an increasedoes not cause undesirable toxicity effects. Patients may be treated fora minimum number of cycles, as disclosed herein. Complete or partialresponse may require additional treatment cycles. Treatment may becontinued as long as the patient continues to benefit.

I. Example 9

The effects of AZA dose and schedule (short-term vs. extended) onpharmacodynamic markers such as DNMT1 depletion, DNA methylation, andDNA damage were evaluated in MDA-MB-231 breast cancer cells in vitro andin vivo. For in vitro experiments, MDA-MB-231 cells were treated dailywith 0.1 or 0.3 μM AZA for up to 12 days, and harvested at various timesduring treatment, as well as up to 12 days following treatment. For invivo studies, MDA-MB-231 tumor-bearing mice were dosed (ip) with 1 or 3mg/kg AZA daily for 3, 7, 14, 21, or 28 days and tumors were harvestedduring and at several time points after the dosing period. DNA and celllysates were prepared (from cell pellets or xenograft tumors) for DNAmethylation analysis (LINE-1 or EpiTech Methyl qPCR assay) andDNMT1/γH2AX western blotting, respectively.

In both in vitro and in vivo studies, AZA caused a rapid (by 8 hourspost in vivo dose), dose-dependent depletion of DNMT1 protein; when AZAtreatment was halted, DNMT1 protein levels returned to basal levelswithin 3-4 days. Consistent with these results, AZA in vitro and in vivocaused a dose-dependent decrease in DNA methylation (LINE-1 andgene-specific) and further reduction in DNA methylation with additionaldays of AZA dosing. In vitro, DNA methylation returned to basal levelsupon AZA removal (within 8 days); the kinetics of DNA re-methylation wasslower in more hypomethylated DNA. Lastly, DNA damage was not observedin tumors from mice until 14 or 21 days of dosing with 3 mg/kg or 1mg/kg AZA, respectively.

Thus, extended AZA dosing maintained low DNMT1 levels and DNAmethylation, and induced DNA damage. These results provide a strongrationale for extended AZA dosing for the treatment of cancer patients.

J. Example 10

In patients treated with 5-azacytidine alone or in combination with anadditional therapeutic agent such as CDBCA or ABX, the pharmacodynamiceffects are determined using one or more methods provided in the tablebelow. In addition, the methods below can be used as predictivebiomarkers to predict clinical response to treatments.

Tissue Analyte Assay PD¹ Pred.² Whole Blood Genomic DNA Infinium ® ✓(PBMCs) Methylation27 Array Plasma Free DNA Infinium ® ✓ ✓Methylation450 Array Tumor Genomic DNA Infinium ® ✓ ✓ (Fresh Frozen)Methylation450 Array RNA Candidate gene ✓ ✓ expression analysis TumorProtein (IHC) DNMT1, DNMT3A, ✓ ✓ (FFPE) pH2AX, cPARP Candidate short ✓ ✓half-life proteins ¹PD: Change at Day 15 from baseline. ²PredictiveBiomarker: Baseline in relation to clinical response.

In one embodiment, PBMC DNA was used for assessing changes in DNAmethylation, using assays such as LINE-1 methylation, %5mdC mass spec,Infinium® Methylation27 Array, and Infinium® Methylation450 Array. Inone study, DNA methylation (LINE-1) in PBMC DNAs from patients inclinical studies described in Example 1, dosed with 200 mg oral AZAalone or in combination with CDBCA or ABX, was measured on Days 1, 8,and 15 of 21-day cycle. Decreases in LINE-1 methylation were observedfor two patients in Arm C. In one study, %5mdC in PBMC DNAs frompatients in clinical studies described in Example 1, dosed with 200 mgoral AZA alone or in combination with CDBCA or ABX, was measured on Days1, 8, and 15 of 21-day cycle. In one study, methylation levels weremeasured on Days 1, 8, and 15 of 21-day cycle using Infinium®Methylation27 Array (patients dosed with 200 mg oral AZA alone or incombination with CDBCA or ABX) and density profiles of averagemethylation levels were analyzed. Upon treatment, decreases inhypermethylated loci (beta>0.7) were observed in PBMCs of Arm Cpatients; no change in DNA methylation was observed in PBMCs of Arm Apatients, and minor decrease in DNA methylation was observed in PBMCs ofArm B patients. In another study, methylation levels were measured usingInfinium® Methylation450 Array (patients dosed with 300 mg oral AZAalone or in combination with CDBCA or ABX) and density profiles ofaverage methylation levels were analyzed, as well as % change ofhypermethylated loci (beta>0.7) upon treatment. The data suggested thatthe decreases in DNA methylation correlated with the PK exposure of AZAin the patients.

In summary, DNA hypomethylation in PBMCs was observed in patients dosedwith 200 mg oral AZA alone or in combination with an additionaltherapeutic agent (5/6 Arm C patients; 2/6 Arm B patients; and 0/6 Arm Apatients). DNA hypomethylation in PBMCs was observed in patients dosedwith 300 mg oral AZA alone or in combination with an additionaltherapeutic agent (3/3 Arm C patients; and 2/4 Arm A patients).Decreases in DNA methylation appeared to correlate with PK exposure ofAZA in the patients, for example, were observed in patients withAUC_(inf)>350 ng*hr/mL.

K. Example 11

FIG. 6 shows in vitro modeling of the dosing schema of the clinicalstudy described in Example 1. DNA hypomethylation (e.g., LINE-1, p16)was measured 72 hours after AZA treatment alone or in combination withCBDCA or ABX. p16 (mRNA) re-expression was determined 72 hours after AZAtreatment alone or in combination with CBDCA or ABX. Ninety-two cancercell lines were tested in order to identify specific tumor types thatbecome sensitized to CBDCA or ABX after treatment with AZA (bladdercancer n=8; head and neck cancer n=8, breast cancer n=21; lung cancern=35; pancreatic cancer n=7; ovarian cancer n=7; and melanoma n=6). Theresult from this study can also be used to identify predictivebiomarkers for enhanced sensitivity to the combination treatment ofCBDCA or ABX with AZA.

Interaction of AZA treatment with ABX treatment was evaluated, in thefollowing cancer cell lines: (1) bladder cancer cell lines, including5637, J82, HT-1376, SCaBER, TCCSUP, and UM-UC-3, which showed additiveeffects; (2) head and neck cancer cell lines, including A253, BHY,CAL-27, CAL-33, and FIN, which showed additive effects; (3) breastcancer cell lines, including ZR-75-1, CAL-51, MDA-MB-231, BT-549,Hs578t, HCC1500, HCC-1187, and ZR-75-30, which showed additive effects;(4) pancreatic cancer cell lines, including MiaPaca-2, which showedsynergistic effects; (5) NSCLC cell lines, including H1792, which showedsynergistic effects; and H460, H1299, H23, H1975, H2122, H838, H28,H1838, CALU-3, H2030, H1437, H596, H647, and H1650, which showedadditive effects; (6) ovarian cancer cell lines, including OVCAR-3,OVCAR-5, OVCAR-8, SKOV3, and IGR-OV1, which showed additive effects; and(7) melanoma cell lines, including Malme 3M and SKMEL5, which showedadditive effects. However, antagonism was observed for combination ofAZA with ABX in ¼ of the cell lines tested, including breast (⅓), NSCLC(¼), and melanoma (⅔) cell lines. In other experiments, antagonism wasalso observed in some cell lines with DAC (decitabine) priming.

Interaction of AZA treatment with CBDCA treatment was evaluated, in thefollowing cancer cell lines: (1) bladder cancer cell lines, includingUM-UC-3, which showed synergistic effects; and 5637, J82, HT-1376,SCaBER, and TCCSUP, which showed additive effects; (2) head and neckcancer cell lines, including Detroit562 and FADU, which showedsynergistic effects; and A253, BHY, CAL-27, CAL-33, HN, and RPMI-2650,which showed additive effects; (3) breast cancer cell lines, includingBT-549, Hs578t, MDA-MB-157, SUM-149, and HCC-38, which showedsynergistic effects; and T47D, ZR-75-1, CAL-51, CAL-120, MCF7, HCC1500,AU565, HCC-1187, MDA-MB-436, and ZR-75-30, which showed additiveeffects; (4) pancreatic cancer cell lines, including BxPC3, MiaPaca-2,and Hs766t, which showed synergistic effects; (5) NSCLC cell lines,including H460, H1299, A549, H838, HOP62, H1792, H1838, H1755, H2030,H1437, HOP92, and H2110, which showed synergistic effects; and H23,H1975, H226, H2122, H28, H2228, H727, SK-LU-1, H520, H596, H647, H1568,H1944, and H1650, which showed additive effects; (6) ovarian cancer celllines, including OVCAR-3, OVCAR-4, OVCAR-5, OVCAR-8, SKOV3, and IGR-OV1,which showed additive effects; and (7) melanoma cell lines, includingSKMEL5, which showed synergistic effects; and M14, Malme 3M, MeWO,SKMEL2, and SKMEL28, which showed additive effects. Additivity orsynergy was observed for combination of AZA with CBDCA in the majorityof cell lines tested (e.g., about ⅓ NSCLC cell lines showed synergy). Inother experiments, additivity or synergy was also observed in some celllines with DAC (decitabine) priming.

Moreover, for the combination of AZA with CBDCA, selected panels of celllines which showed synergistic or additive effects were further studiedin order to identify predictive biomarkers to predict synergy of thecombination. The selected cell lines included (1) UM-UC-3 (bladder),FADU (head and neck), MiaPaca-2 (pancreatic), H838 (NSCLC), H2110(NSCLC), and HOP62 (NSCLC), which showed synergistic effects, and (2)CAL-120 (breast), AU565 (breast), Detroit562 (head and neck), H520(NSCLC), H1838 (NSCLC), H1568 (NSCLC), and CALU-6 (NSCLC). Basal geneexpression and DNA methylation were compared. In addition, AZA-inducedchanges in gene expression and DNA methylation were compared. The extentof synergy was calculated using AAUC values. Strong synergy was observedin 18 hours to 72 hours or more of AZA priming, for example, in HOP62,UM-UC-3, and FADU.

Similar results were observed in the kinetics of AZA-induced DNMT1depletion when comparing synergistic cell lines with additive cell lines(e.g., HOP62 vs. H1568; H838 vs. H520, FADU vs. Detroit562), and similarextent of DNMT1 depletion at 48 hours were observed when comparingsynergistic cell lines with additive cell lines. Similar effects of DNAhypomethylation (LINE-1) was observed in when comparing synergistic celllines with additive cell lines upon AZA treatment alone or incombination with CBDCA (e.g., in FADU, Detroit562, HOP62, and H520).Dramatic increase in PARP cleavage in the synergistic cell line H838 wasobserved upon treatment with combination of AZA and CBDCA, suggestingsynergistic effect of the combination on PARP cleavage in H838 cells.

Furthermore, to identify predictive biomarkers for enhanced sensitivity,basal gene expression, promoter methylation, and mutation status areanalyzed in selected cancer cell lines. Moreover, gene expression andpromoter methylation changes upon AZA treatment are analyzed in selectedcancer cell lines.

L. Example 12 Clinical Evaluation of Anti-Tumor Activity of5-Azacytidine

In one study, eligible subjects were stratified according to specifictumor types, who were treated with 5-azacytidine alone at 300 mg/day for14 consecutive days in a 21-day cycle until progression or intolerabletoxicity. All subjects were evaluated for safety, pharmacokinetics, andtumor response per RECIST 1.1. Subjects with tumors (7 patients withNPC; 4 with cervical carcinoma; 3 with Merkel cell tumors; and 4 withHNSCC) were treated and evaluated. The median number of prior regimensfor this cohort was 3 (range 1-6). The median number of 5-azacytidinecycles administered was 4 (range 1-16). Treatment was well tolerated.Grade 3-4 adverse events reported in >10% of study subjects includedneutropenia (36.8%), anemia (10.5%), hyponatremia (10.5%) and asthenia(10.5%). Four subjects, three with NPC and one with cervical carcinoma,achieved an objective response. Duration of responses ranged from atleast 36 days to 192 days. An additional subject achieved a bestresponse of stable disease lasting >180 days. The disease control rateamong patients with NPC was particularly noteworthy with 6/7 subjectsachieving a response of stable disease (SD) or better and 3 subjectswere still receiving 5-azacytidine treatment after 64, 203, and 240days.

M. Example 13 Evaluation of Anti-Lesion Activity of 5-Azacytidine

In one study, eligible subjects with nasopharyngeal carcinoma weretreated with 5-azacytidine. As shown in FIG. 9, lesions decreased insize in response to treatment with 5-azacytidine.

N. Example 14 Clinical Evaluation of Anti-Tumor Activity of5-Azacytidine

Clinical studies are conducted to assess the the safety and efficacy ofsingle agent 5-azacytidine (oral azacitidine) in previously treatedsubjects with locally advanced or metastatic nasopharyngeal carcinoma,after having received one to two previous regimens, including a platinumagent. Approximately 50-55 subjects will be enrolled according to aSimon two-stage design; if the predefined activity is met (4 objectiveresponses out of the first 18 subjects) then will continue to treat upto 50 subjects. Subjects will be treated until radiologic diseaseprogression, or unless otherwise specified. Tumor response will beassessed according to RECIST 1.1 initially every 6 weeks (+/−5 days) andthen until documented disease progression, start of new anticancertherapy, or withdrawal of consent. All subjects will also be followedfor survival and subsequent anticancer therapies.

The anticipated duration of the study is 8 months for enrollment ofsubjects and 4 months for follow up from the last subject first visit.The entire study duration is anticipated to be approximately 12 months.

All subjects will be followed for 28 days after discontinuing treatmentfor safety and monitoring of adverse events, for response untilprogression (if applicable), new anticancer therapies and for survivaluntil the trial is mature for analysis, or the End of Trial, whicheveroccurs earlier.

The End of Trial is defined as either the date of the last visit of thelast subject to complete the study, or the date of receipt of the lastdata point from the last subject that is required for primary, secondaryand/or exploratory analysis, as pre-specified in the protocol and/or theStatistical Analysis Plan, whichever is the later date.

Screening evaluations will be performed for all subjects to determinestudy eligibility. These evaluations may be completed within 21 days offirst dosing unless noted below. Safety laboratory analyses and allassessments will be performed locally. Screening laboratory values mustdemonstrate subject eligibility, but may be repeated within thescreening window if necessary. The following will be performed atscreening as specified in the Table of Events, after informed consenthas been obtained:

-   -   Cancer history (including specific information regarding        diagnosis, staging, histology)    -   Demographics (initials, date of birth, sex, race, and        ethnicity-if allowed by local regulations)    -   Prior cancer therapies includes surgery, radiation, systemic or        any other therapy for the subject's cancer    -   Complete medical history (all relevant medical conditions        occurring ≧28 days before screening should also be included)    -   Prior and concomitant procedures (including all procedures        occurring ≦28 days before screening)    -   Prior and concomitant medication evaluation (including those        taken ≦28 days before screening, except for those taken for        cancer)    -   Physical examination (source documented only), height, weight    -   Vital signs (including blood pressure, temperature, and heart        rate)    -   Eastern Cooperative Oncology Group (ECOG) Performance status    -   12-lead electrocardiogram (ECG)    -   Response assessment/tumor evaluation. Subjects with historical        tumor scans evaluable per RECIST 1.1 performed ≦28 days before        the first dose need not repeat scans for the purposes of        screening    -   Complete blood count (CBC) with differential, including but not        limited to red blood cell (RBC) count, hemoglobin, hematocrit,        white blood cell (WBC) count, absolute neutrophil count (ANC),        and platelet count. ANC should be measured with automated count        where available    -   Chemistry panel including, but not limited to, sodium,        potassium, calcium, chloride, blood urea nitrogen (BUN),        creatinine, glucose, albumin, total protein, alkaline        phosphatase, bilirubin (total and direct), aspartate        aminotransferase/serum glutamic oxaloacetic transaminase        (AST/SGOT), alanine aminotransferase (ALT/SGPT)    -   Coagulation tests including, prothrombin time (PT), partial        thromboplastin time (PTT), international normalized ratio (INR).    -   Urinalysis (a urine dipstick may be used)    -   Pregnancy test is required for all female subjects of        childbearing potential. Serum β-hCG pregnancy test will be        performed at screening. Urine pregnancy test will be performed        to assess subject eligibility within 72 hours prior to the first        administration of IP, if the serum pregnancy test did not        already occur with 72 hours of dosing (negative results required        for IP administration).    -   Adverse event assessment begins when the subject signs the        informed consent form

The subject will begin treatment upon confirmation of eligibility. Thesubject must start treatment within 21 days of signing the informedconsent form (ICF). For all subsequent visits, an administrative windowof ±2 days is permitted. Treatment cycles are 21 days in duration. Thefollowing evaluations will be performed at the frequency specified inthe Table of Events. The evaluations should be performed prior to dosingon the visit day, unless otherwise specified:

-   -   Concomitant medications evaluation    -   Concomitant procedures evaluation    -   Physical examination (source documented only)    -   Vital signs: on-treatment vital sign measurements will be source        documented only. However, if an abnormal (out of range) value is        reported at any given visit, that parameter should be collected        in the CRF at every subsequent scheduled visit until it returns        to normal, and as an AE if appropriate.    -   Weight    -   Complete blood count with differential    -   Chemistry panel    -   PK sample    -   Tumor biopsy    -   Body surface area calculation (source document only)    -   ECOG Performance status    -   Adverse event evaluation (continuously)    -   Response assessment/tumor evaluation    -   Urine pregnancy test (prior to dosing on Day 1)

An end of treatment (EoT) evaluation should be performed for subjectswho are withdrawn from treatment for any reason as soon as possibleafter the decision to permanently discontinue treatment has been made.The following evaluations will be performed as specified in the Table ofEvents:

-   -   Physical examination    -   Vital signs    -   Concomitant medications evaluation    -   Concomitant procedures evaluation    -   ECOG performance status    -   Adverse event evaluation (monitored through 28 days after the        last dose of IP)    -   Complete blood count (with differential)    -   Chemistry panel    -   Coagulation tests    -   Urine β-hCG level for females of childbearing potential    -   Response assessment/tumor evaluation will be continued at the        schedule defined in the Table of Events, and does not need to be        performed specifically for the EOT visit except as specified        otherwise    -   12-lead ECG

All subjects who discontinue treatment for reasons other than diseaseprogression, start of new anticancer therapy, or withdrawal of consentfrom the entire study will be followed for tumor response assessmentsand subsequent anticancer therapies as specified.

After the EOT visit, all subjects will be followed every 8 weeks forsurvival until the trial is the End of Trial. Subsequent anticancertherapies should be collected at the same schedule. New anticancertherapy includes (but is not limited to) any systemic or localmedication, surgery, radiation, or any other therapy intended to treatthe subject's cancer. Survival follow-up may be conducted by recordreview (including public records) and/or telephone contact with thesubject, family, or the subject's treating physician.

Response assessments (tumor evaluations) should be performed atscreening within 28 days before the start of IP, and every 6 weeks (±5days) from Cycle 1 Day 1 for the first two tumor evaluations and thenevery 9 weeks until disease progression, start of a new anticancertherapy, or withdrawal of consent from the entire study. Subjects withhistorical tumor scans evaluable per RECIST 1.1 performed ≦28 daysbefore the first dose need not repeat scans for the purposes ofscreening. Evaluation of response should be performed using RECIST 1.1guidelines, by investigator assessment. If necessary, an independent(central) assessment of response may be conducted by the sponsor afterstudy completion.

Response will be assessed using RECIST 1.1, according to investigatorassessment. Response assessments include CT scan or MRI of the head andneck with supraclavicular node imaging, the chest and abdomen/pelvis,neurological examination with facial nerve evaluation, and bone scans atbaseline for all subjects. Bone scans will be repeated only if thesubject is symptomatic or with known bone metastasis. The same mode ofimaging for lesion evaluation at screening must be used consistentlythroughout the study. Adherence to the planned imaging schedule iscritical regardless of dose delays or unscheduled or missed assessments.The CT imaging should include contrast unless medically contraindicated.Conventional CT should be performed with contiguous cuts of 5 mm or lessin slice thickness. Spiral CT should be performed by use of a 5 mmcontiguous reconstruction algorithm. All subjects with evidence ofobjective tumor response (CR or PR) should have the response confirmedwith repeat assessments at the next scheduled scan, but after no lessthan 4 weeks. Response assessments must have occurred ≧6 weeks fromCycle 1 Day 1 to be considered as SD for a best response.

Subjects who are symptomatic for brain metastasis at screening mustundergo a brain scan to confirm eligibility. All subjects must undergobone scans at baseline.

Following confirmation of eligibility during the screening phase, aselected number of sites and a subset of enrolled subjects willparticipate in the PK sample collection. Six subjects of Asian-Pacificethnicity initially being administered 200 mg 5-azacytidine on Days 1-14of a 21-day cycle will be asked to participate in PK sample collectionat these selected sites capable of participating in obtaining in thisdata. An additional 6 subjects of Asian-Pacific ethnicity willparticipate in the PK sampling if the dose is escalated to 300 mg inthis population. On each PK day (i.e., Cycle 1 Day 1 and 14), subjectswill ingest IP in the clinic after performing the required overnightfasting, taking mandatory antiemetic premedication (eg, odansetron), andcompleting the required pre-dose assessments (where applicable), witheach dose being given at approximately the same time of day. The exactdate and time of dosing will be recorded in the source documents andappropriate CRF.

Blood samples for azacitidine PK assessment will be collected prior toeach dose administration (pre-dose) and over the 8-hour period followingeach dose administration (0.25, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 6, and 8hours post-dose or similar schedule).

After an overnight fast of at least 8 hours and after performance of allrequired pre-dose assessments and procedures (including obtaining thepre-dose PK sample), subjects should ingest antiemetic premedication(eg, odansetron) with 240 mL of water. Approximately 30 minutes latersubjects will ingest 5-azacytidine with 240 mL of room temperaturewater. No food will be allowed for at least 2 hours post-dose. Water canbe allowed as desired except for 1 hour before and after 5-azacytidineadministration. The only water permitted in the 1 hour period before5-azacytidine administration is the 240 mL of water for antiemeticingestion.

Predictive biomarkers may allow prospective identification of patientswho are likely to benefit clinically from 5-azacytidine as a singleagent. Although the predictive diagnostic analyses in the current trialare exploratory in nature, they could reveal associations betweenbiomarkers and responses that could provide a basis for futurediagnostically driven studies.

Approximately 50-55 subjects with advanced or metastatic NPC will beenrolled. The study will be conducted at sites globally.

Subjects must satisfy the following criteria to be enrolled in thestudy:

-   -   1. Subject is ≧18 years of age or older the time of signing the        informed consent document.    -   2. Subject has a histological or cytological diagnosis of        undifferentiated or poorly differentiated nasopharyngeal        carcinoma that is locally advanced or metastatic.    -   3. Subject has failed one to two prior lines of treatment.    -   4. Subject has received a platinum containing regimen.    -   5. Subject has an ECOG performance status 0-2.    -   6. Subject has radiographically-documented measurable disease,        as per RECIST 1.1.    -   7. Subject has adequate organ functions, evidenced by the        following:        -   a. AST (SGOT), ALT (SGPT)≦2.5× upper limit of normal range            (ULN), or ≦5× upper limit of normal range if liver            metastasis present        -   b. Total bilirubin≦1.5×ULN        -   c. Creatinine≦1.5×ULN        -   d. Potassium within normal range, or correctable with            supplements    -   8. Subject has adequate bone marrow function, evidenced by the        following:        -   e. Absolute neutrophil count ≧1.5×10⁹ cells/L        -   f. Platelets≧100×10⁹ cells/L        -   g. Hemoglobin≧9 g/dL    -   9. Females of childbearing potential (FCBP) (defined as a        sexually mature woman who 1) has not undergone a hysterectomy        (the surgical removal of the uterus) or bilateral oophorectomy        (the surgical removal of both ovaries) or, 2) has not been        naturally postmenopausal for at least 24 consecutive months (ie,        has had menses at any time during the preceding 24 consecutive        months) must:        -   h. Have two negative pregnancy tests as verified by the            study doctor prior to starting study therapy: negative serum            pregnant test, sensitivity of at least 25 mIU/mL at            screening and have a negative serum or urine pregnancy test            within 72 hours prior to starting study therapy (note that            the screening serum pregnancy test can be used as the test            prior to starting study therapy in the treatment phase if it            is performed within the 72-hour timeframe). She must agree            to ongoing pregnancy testing during the course of the study,            and after end of study therapy. This applies even if the            subject practices true abstinence from heterosexual contact.        -   i. Either commit to true abstinence from heterosexual            contact (which must be reviewed on a monthly basis) or agree            to use, and be able to comply with, effective contraception            (oral, injectable, or implantable hormonal contraceptive;            tubal ligation; intra-uterine device; barrier contraceptive            with spermicide; or vasectomized partner) without            interruption, 28 days prior to starting study drug, during            the study therapy (including dose interruptions), and for 3            months after discontinuation of 5-azacytidine.    -   10. Male subjects with a female partner of childbearing        potential must agree to the use of a physician-approved        contraceptive method throughout the course of the study and        avoid fathering a child during the course of the study and for 6        months following the last dose of 5-azacytidine.    -   11. Subject understands and voluntarily signs an ICF prior to        any study related assessments/procedures are conducted.    -   12. Subject is able to adhere to the study visit schedule and        other protocol requirements. True abstinence is acceptable when        this is in line with the preferred and usual lifestyle of the        patient. [Periodic abstinence (eg, calendar, ovulation,        symptothermal, post-ovulation methods) and withdrawal are not        acceptable methods of contraception].

The presence of any of the following will exclude a subject fromenrollment:

-   -   1. Subject has a history of, or current brain metastasis. In        subjects who are symptomatic, a brain scan is required to        exclude metastasis. Primary NPC tumors that are        touching/penetrating the skull are not considered brain        metastasis.    -   2. Subject has any other malignancy within 5 years prior to        randomization, with the exception of adequately treated in-situ        carcinoma of the cervix, uteri, or non-melanomatous skin cancer        (all treatment of which should have been completed 6 months        prior to enrollment).    -   3. Subject has been previously treated with azacitidine (any        formulation), decitabine or any other hypomethylating agent    -   4. Subject has a history of inflammatory bowel disease (e.g.,        Crohn's disease, ulcerative colitis), celiac disease (i.e.,        sprue), prior gastrectomy or upper bowel removal, or any other        gastrointestinal disorder or defect that would interfere with        the absorption, distribution, metabolism or excretion of the        study drug and/or predispose the subject to an increased risk of        gastrointestinal toxicity.    -   5. Subject has an impaired ability to swallow oral medication.    -   6. Subject has persistent diarrhea or malabsorption ≧NCI CTCAE        grade 2, despite medical management.    -   7. Subject has significant active cardiac disease within the        previous 6 months including unstable angina or angina requiring        surgical or medical intervention, significant cardiac        arrhythmia, or New York Heart Association (NYHA) class 3 or 4        congestive heart failure.    -   8. Subject has a known or suspected hypersensitivity to        Azacitidine, mannitol, or any other ingredient used in the        manufacture of 5-azacytidine.    -   9. Subject has a known history or current diagnosis of Human        Immunodeficiency Virus (HIV) infection, regardless of treatment        status.    -   10. Subject has any other concurrent severe and/or uncontrolled        medical condition that would, in the investigator's judgment,        contraindicate patient participation in the clinical study (eg,        chronic pancreatitis, chronic active hepatitis, etc.).    -   11. Subject has had major surgery within 14 days prior to        starting study drug or has not recovered from major side        effects.    -   12. Subject has received another investigational therapy within        28 days or 5 half lives of randomization/enrollment, whichever        is shorter.    -   13. Subject has not recovered from the acute toxic effects of        prior anticancer therapy, radiation or major surgery/significant        trauma.    -   14. Subject has had radiotherapy ≦4 weeks or limited field        ration for palliation ≦2 weeks prior to starting study drug,        and/or from whom ≧30% of the bone marrow was irradiated.        -   Prior radiation therapy to a target lesion is permitted only            if there has been clear progression of the lesion since            radiation was completed.    -   15. Subject is pregnant or breast feeding.    -   16. Subject has any significant medical condition, laboratory        abnormality, or psychiatric illness that would prevent the        subject from participating in the study.    -   17. Subject has any condition including the presence of        laboratory abnormalities, which places the subject at        unacceptable risk if he/she were to participate in the study.    -   18. Subject has any condition that confounds the ability to        interpret data from the study.

Celgene Corporation will supply 5-azacytidine 100-, 150-, and/or 200-mgtablets for oral administration. All tablets will be packaged in blistercards. Each tablet is formulated using excipients that are generallyregarded as safe and are used in marketed drug products. A list ofexcipients included in the formulations is provided in the azacitidineIB.

All IP must be stored in an area free of environmental extremes and asecured area to prevent unauthorized access, as directed on the packagelabel. A temperature log must be maintained.

5-Azacytidine will be administered at a dose of 300 mg orally daily for14 days of a 21-day cycle. The first 6 Asian subjects will receive5-azacytidine at a dose of 200 mg orally daily for 14 of 21 days ofCycle 1; and if well-tolerated, all others subjects will be administeredat the 300 mg daily dose for 14 days of a 21-day cycle. Anti-emeticmedication (not supplied by the sponsor) may be taken 30 minutes priorto IP administration at the investigator's discretion.

Subjects will ingest IP with approximately 240 mL (8 ounces) of roomtemperature water. Investigational product may be taken on an emptystomach or with food (a light breakfast or meal of up to approximately600 calories), except on PK sampling days.

A maximum of 2 dose reductions will be allowed from the original doseExcept for those patients starting at 200 mg, only one dose reduction ispermitted. 5-Azacytidine may be withheld for up to 7 days between theend of one Cycle and the start of the next Cycle to allow hematologiccriteria to recover sufficiently for the next Cycle to begin. Themaximum number of days that a dose may be withheld without requiring adose reduction is 7 days. The maximum number of days that a dose may bewithheld due to unacceptable toxicity before a subject is permanentlydiscontinued from the study is 14 days. For the purposes of doseadjustments, unacceptable toxicity will be defined as any AE that isdeemed by the investigator to be related to 5-azacytidine that poses amedical risk or substantial discomfort to the subject including but notlimited to Grade 3 or 4 hematologic or non-hematologic toxicity.

All efforts should be made to administer IP on all of the scheduled daysof each 21-day treatment cycle. Any missed doses during that periodshould not be added after the last scheduled day of administration, butshould be returned by the subject for IP accountability.

Overdose, as defined for this protocol, refers to 5-azacytidine only. Ona per dose basis, an overdose is defined as the following amount overthe protocol-specified dose of 5-azacytidine assigned to a givensubject, regardless of any associated adverse events or sequelae.

-   -   PO—any amount over the protocol-specified dose        On a schedule or frequency basis, an overdose is defined as        anything more frequent than the protocol required schedule or        frequency. Complete data about drug administration, including        any overdose, regardless of whether the overdose was accidental        or intentional, should be reported in the case report form. For        a drug to be subject to the overdose definition it must be both        a required drug (for which dosing information is collected on        the CRF), and either a control (active or placebo) or        experimental drug or regimen that is directly under study.

The following events are considered sufficient reasons for discontinuinga subject from the investigational product:

-   -   Adverse Event    -   Progressive disease    -   Symptomatic deterioration (global deterioration of health        status)    -   Physician decision    -   Withdrawal by subject    -   Death    -   Lost to follow-up    -   Protocol violation    -   Other (to be specified on CRF)

The reason for discontinuation should be recorded in the CRF and in thesource documents. The decision to discontinue a subject remains theresponsibility of the treating physician, which will not be delayed orrefused by the sponsor. However, prior to discontinuing a subject, theinvestigator may contact the Medical Monitor and forward appropriatesupporting documents for review and discussion.

The following events are considered sufficient reasons for discontinuinga subject from the study follow-up periods:

-   -   Withdrawal of consent    -   Death    -   Lost to follow up    -   Protocol violation    -   Other

The reason for discontinuation should be recorded in the CRF and in thesource documents.

Efficacy analyses will be performed on the intent-to-treat (ITT)population that includes all subjects enrolled. The hypothesis testingfor the ORR will be based on the exact binomial distribution. The pointestimate and 95% confidence interval (CI) of ORR will be provided. TheKaplan-Meier procedure will be used to estimate the median PFS and 95%CI. Subjects who die, regardless of the cause of death, will beconsidered to have had an event. Subjects who withdraw consent for thestudy will be considered censored at the time of withdrawal. Subjectswho are still alive at the time of the clinical data cut-off date willbe censored. All subjects who were lost to follow-up prior to theclinical data cut-off date will also be considered censored at the timeof last contact. The Kaplan-Meier procedure will be used to estimateproportions of event-free interval for OS. Medians and their two-sided95% confidence intervals will be provided.

The safety population will be included for safety analysis. Safety andtolerability will be monitored through continuous reporting of adverseevents and serious adverse events, laboratory abnormalities, andincidence of patients experiencing dose modifications, doseinterruptions, and/or premature discontinuation of study drug. Data fromall subjects who receive one or more doses of study drug will beincluded in the safety analyses. Adverse events, physical examinations(including vital sign measurements), clinical laboratory information,and concomitant medications/procedures will be tabulated and summarized.

Adverse events will be analyzed in terms of TEAE defined to be any eventthat begins or worsens in grade after the start of study drug through 28days after the last dose of IP. Adverse events will be summarized byseverity/grade based on the CTCAE Version 4.0 and relationship to studytreatment. If a subject experiences the same AE multiple times duringthe treatment, the event will be counted only once and by the greatestseverity. AEs, as well as treatment emergent AEs, will be summarized bysystem organ class, and preferred term. Adverse events leading to deathor to discontinuation from treatment, events classified as CTCAE Grade 3or Grade 4, IP related events, and serious adverse events will besummarized separately. Adverse events will be coded according to theMedical Dictionary for Regulatory Affairs (MedDRA). Adverse events ofspecial interest of 5-azacytidine identified in previous trials will besummarized in a similar manner.

Pharmacokinetic measures are incorporated into the study to assess theextent of exposure, and to explore the relationship between5-azacytidine and PD effects. Blood samples for PK will be collectedfrom a subset of subjects at selected visits. As appropriate, thefollowing PK parameters, but not limited to, will be determined fromplasma concentration versus time profile data:

-   -   Area under the plasma concentration time-curve (AUC);    -   Peak (maximum) plasma concentration (C_(max));    -   Terminal half-life (t_(1/2));    -   Time to maximum plasma concentration (T_(max));    -   Clearance (apparent, CL/F); and,    -   Volume of distribution (apparent, Vz/F).        Descriptive statistics (N, mean, SD, coefficient of variation        [CV %], geometric mean, geometric CV %, median, min, and max)        will be provided for all data. Results will be presented in        tabular and graphic forms as appropriate. Sample collection kits        and detailed instructions for PK sample collection, processing,        storage, shipping and handling will be provided to the        investigator sites upon study initiation. Plasma azacitidine        will be measured using validated liquid chromatography-mass        spectrometry methods (LC-MS/MS).

The present disclosure has been described in connection with certainembodiments and examples; however, unless otherwise indicated, theclaimed invention should not be unduly limited to such specificembodiments and examples.

What is claimed is:
 1. A method for treating a subject having a cancer,wherein the method comprises orally administering to the subject apharmaceutical composition comprising 5-azacytidine, or apharmaceutically acceptable salt, solvate, or hydrate thereof; and themethod optionally further comprises administering at least oneadditional therapeutic agent.
 2. The method of claim 1, wherein thecancer is a relapsed or refractory solid tumor.
 3. The method of claim1, wherein the cancer is nasopharyngeal carcinoma, cervical carcinoma,cervical adenocarcinoma, non-small cell adenocarcinoma of the lung,colon Kras adenocarcinoma, or colon adenocarcinoma.
 4. The method ofclaim 1, wherein the additional therapeutic agent is an anti-canceragent.
 5. The method of claim 4, wherein the additional therapeuticagent is a platinum agent.
 6. The method of claim 5, wherein theadditional therapeutic agent is carboplatin.
 7. The method of claim 4,wherein the additional therapeutic agent is a taxane.
 8. The method ofclaim 7, wherein the additional therapeutic agent is paclitaxel.
 9. Themethod of claim 7, wherein the additional therapeutic agent ispaclitaxel protein-bound particles.
 10. The method of claim 7, whereinthe additional therapeutic agent is Abraxane.
 11. The method of claim 1,wherein the method comprises the steps of: (i) administering5-azacytidine to the subject for 1, 2, 3, 4, 5, 6, or 7 days; and (ii)administering the additional therapeutic agent to the subject for one ormore days.
 12. The method of claim 11, wherein the additionaltherapeutic agent is administered parenterally.
 13. The method of claim11, wherein the additional therapeutic agent is administered orally. 14.The method of claim 11, wherein step (ii) further comprisesadministering 5-azacytidine orally for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, 12, 13, or 14 days.
 15. The method of claim 11, wherein the methodcomprises the sequential steps of: (i) administering 5-azacytidine tothe subject for 7 days; (ii) administering the additional therapeuticagent to the subject for 1 day; (iii) administering 5-azacytidine to thesubject for 6 days; and (iv) repeating steps (i) to (iii) after 7 daysof resting period.
 16. The method of claim 11, wherein the methodcomprises the sequential steps of: (i) administering 5-azacytidine tothe subject for 7 days; (ii) administering 5-azacytidine and theadditional therapeutic agent to the subject for 1 day; (iii)administering 5-azacytidine to the subject for 6 days; and (iv)repeating steps (i) to (iii) after 7 days of resting period.
 17. Themethod of claim 11, wherein the method comprises the sequential stepsof: (i) administering 5-azacytidine to the subject for 7 days; (ii)administering 5-azacytidine and the additional therapeutic agent to thesubject for 1 day; (iii) administering 5-azacytidine to the subject for6 days; (iv) administering the additional therapeutic agent to thesubject for 1 day; and (v) repeating steps (i) to (iv) after 6 days ofresting period.
 18. The method of claim 11, wherein the method comprisesthe sequential steps of: (i) administering 5-azacytidine to the subjectfor 7 days; (ii) administering 5-azacytidine and the additionaltherapeutic agent to the subject for 1 day; (iii) administering5-azacytidine to the subject for 6 days; (iv) administering theadditional therapeutic agent to the subject for 1 day; (v) administering5-azacytidine and the additional therapeutic agent to the subject for 1day after 6 days of resting period; (vi) administering 5-azacytidine tothe subject for 6 days; and (vii) repeating steps (ii) to (vi).
 19. Themethod of claim 1, wherein the method comprises administering5-azacytidine to the subject for at least 1, at least 2, at least 3, atleast 4, at least 5, at least 6, or at least 7 days, beforeadministering the additional therapeutic agent to the subject.
 20. Themethod of claim 19, wherein the additional therapeutic agent isadministered parenterally for one or more days.
 21. The method of claim19, wherein the additional therapeutic agent is administered orally forone or more days.
 22. The method of claim 1, wherein the pharmaceuticalcomposition comprising 5-azacytidine is a tablet.
 23. The method ofclaim 1, wherein the pharmaceutical composition comprising 5-azacytidineis a capsule.
 24. The method of claim 1, wherein the pharmaceuticalcomposition comprises about 100 mg, about 200 mg, about 300 mg, about400 mg, about 500 mg, or about 600 mg of 5-azacytidine.
 25. The methodof claim 1, wherein the pharmaceutical composition comprises about 200mg of 5-azacytidine.
 26. The method of claim 1, wherein thepharmaceutical composition comprises about 300 mg of 5-azacytidine. 27.The method of claim 1, wherein the subject is a human.